A-769662

A-769662 is a reversible AMPK activator with an EC50 value of 0.8 μM. A-769662 exhibits biological activities including metabolic regulation, antitumor effects, and anti-inflammatory properties. A-769662 can be used in metabolic and oncology drug research and development.

AMPK:0.8 μM(EC50)

Methods: Mouse BMSCs were treated with 10 μM A-769662 for 6 days. Western blot analysis was performed at designated time points to detect p-AMPKα (Thr172) and p-ACC (Ser79).
Results: A-769662 significantly increased the phosphorylation levels of AMPKα and its substrate ACC, confirming AMPK signaling activation. [1]

Methods: Male BALB/c mice received intraperitoneal injections of LPS (2 mg/kg). Four hours later, they received intraperitoneal injections of A-769662 (10 mg/kg). Mice were euthanized 8 hours later.
Results: A-769662 significantly reduced TLR-4 mRNA, MyD88 protein, and TNF-α/IL-6 levels while increasing p-AMPK and p-ACC levels.[2]

96-well AMPK assay: AMPK activity is measured by monitoring phosphorylation of the SAMS peptide substrate (20 mM in standard assays and 100 mM in additivity assays) following a previously described protocol (Anderson et al., 2004). To determine whether A-769662-induced AMPK activation occurs in a reversible manner, AMP or A-769662 are preincubated with rat liver AMPK for 10 minutes at 20 times standard assay concentrations prior to dilution and measurement of AMPK activity.

Cell viability of MEF cells treated or not with A-769662 is performed as follows: cells are harvested by trypsinization and incubated with 0.5 mg/mL RNase and 50 μg/mL propidium iodine at room temperature in the dark; cell viability is analyzed by flow cytometry using a FACScanto flow cytometer, using an excitation laser at 488 nm and a propidium iodine fluorescence detection at 600 nm. To determine the proportion of cells in each phase of the cell cycle, cells are harvested by trypsinization, collected by centrifugation, washed in PBS and fixed overnight in 80% ethanol at -20 °C. Subsequently, these fixed cells are centrifuged to remove the fixative and incubated for 20 minutes in the dark at room temperature in PBS containing 0.5 mg/mL RNase and 50 μg/mL propidium iodine. Flow cytometry analysis is performed as above. The proportion of cells in G1, S, and G2 is determined using the MODFIT program. Cell culture pictures are taken at the indicated times using a camera coupled to an inverted microscope with a 20 × objective. (Only for Reference)

Ethanol: 3.61 mg/mL (10.02 mM), Sonication is recommended.

DMSO: 51 mg/mL (141.51 mM), Sonication is recommended.

10% DMSO+40% PEG300+5% Tween 80+45% Saline: 4.5 mg/mL (12.49 mM), Solution.