AMP-activated protein kinase (AMPK) is a conserved serine/threonine kinase that coordinates numerous metabolic processes to maintain cellular energy homeostasis. Given this central role, it is implicated in various metabolic diseases. Pharmacological AMPK activators have been extensively studied with several potent specific compounds developed. In contrast, available inhibitors widely used in the AMPK field which include Compound C and SBI-0206965 are far from selective. Recently, BAY-3827 has emerged as an attractive compound, however its selectivity and structural-functional data available are very limited. We aim to characterise BAY-3827 and illuminate its action mode using multifaceted approaches.
We assessed kinase specificity across a panel of 140 human kinases where mean activity (%) was measured in vitro following 1 uM BAY-3827 treatment. Recorded values (%) revealed compound specificity towards AMPK (1%), and among the top inhibited kinases were RSK1 (5%), MSK1 (6%) and AMPK-related NUAK1 (14%). To characterise the binding mode of BAY-3827, in silico docking simulations of AMPKa and BAY-3827 were performed in Maestro. The best-scoring poses were selected, and together point to a type-I binding mode, with BAY-3827 interacting with hinge residue Val96 and gatekeeper Met93. We incubated isolated mouse skeletal muscle with increasing doses of BAY-3827 ex vivo and observed that glucose uptake induced by an AMPK activator (MK-8722, 10 uM) was significantly reduced at 1 uM (accepted at *p <0.05).
Collectively, these data depict BAY-3827 as an attractive compound with high specificity towards AMPK, serving as a starting point for further characterisation of BAY-3827 in AMPK-related metabolic studies.