O304 activates AMPK and increases cellular ATP
Using a cellular screen, designed to identify compounds which activates AMPK without causing ATP depletion, Betagenon AB has identified a series of small molecule AMPK activator compounds. After optimization, the compound O304 was selected as the candidate drug for further development. O304 increases the levels of p-T172 AMPK in a dose-dependent manner in cells from a variety of species. Moreover, under these conditions, and unlike Metformin, O304 increases cellular ATP.

O304 suppresses the dephosphorylation of p-T172 AMPK
The rate of dephosphorylation of p-T172 AMPKα by the protein phosphatases PP2C and PP1 is a key regulatory step of AMPK activation. Using purified human recombinant enzymes, O304 suppresses the dephosphorylation of p-T172 AMPK trimer by both PP2C and PP1, without inhibiting their activities. O304 also suppresses the dephosphorylation of partially purified rat liver AMPK by PP2C. Like ADP, but unlike AMP, O304 does not allosterically activate AMPK heterotrimers in vitro.