AMPK is a structurally complex energy sensor allosterically regulated by AMP and ADP. In the presence of energetic stress, AMPK up- and down-regulates catabolic and anabolic pathways, respectively, by phosphorylating targets that mediate cytoplasmic signaling and proteins that help regulate gene expression. For years, researchers specializing in cancer and type II diabetes have advocated for the discovery of selective AMPK drugs. To date, however, there are no direct AMPK modulators available for patients. To identify selective AMPK drugs, we first published a high-throughput, fluorescence-based assay biased toward the detection of molecules that bind the regulatory region of AMPK. This assay design was based on the premise that molecules that bind the AMPK regulatory region may be less likely to bind the conserved kinase active site found throughout the kinome. One of the primary hits identified by this assay became the parent molecule for a panel of structural analogs that dose-dependently inhibit binding of a fluorescent probe to the AMPK regulatory region. These analogs have now been characterized in orthogonal protein-based and cell-based assays.