In vivo, C57BKS db/db mice are administered a 15 mg/kg/day dose of ZLN024 by daily gavage. After 4 weeks of treatment, ZLN024 improves glucose tolerance. ZLN024 reduces the fasting blood glucose by 15%. Liver tissue weight, triacylglycerol and the total cholesterol content are decreased^[1].

ZLN024 effectively stimulates both active AMPK heterotrimers and inactive α1 subunit truncations α1 (1-394) and α1 (1-335), although it does not affect α1 (1-312). For ZLN024 to activate AMPK, Thr-172 must be pre-phosphorylated by an upstream kinase, and ZLN024 also shields AMPK Thr-172 from dephosphorylation by PP2Cα. In L6 myotubes, ZLN024 activation of AMPK leads to increased glucose uptake and fatty acid oxidation without altering the ADP/ATP ratio. Identified through random screening using a scintillation proximity assay (SPA) targeting the AMPK α1β1γ1 heterotrimer, ZLN024 has been pinpointed as a novel AMPK activator. It activates AMPK α1β1γ1 and its homologue α2β1γ1 in a dose-dependent manner, enhancing the activity of α1β1γ1 by 1.5-fold with an EC50 of 0.42 µM, and that of α2β1γ1 by 1.7-fold with an EC50 of 0.95 µM. Additionally, ZLN024 boosts the activity of recombinant AMPK α1β2γ1 and α2β2γ1 by 1.7-fold and 1.6-fold, with EC50 values of 1.1 µM and 0.13 µM, respectively^[1].