2-[2-(4-(trifluoromethyl)phenylamino)thiazol-4-yl]acetic acid (Activator-3) is a potent activator of AMPK

Abstract

AMPK is considered as a potential high value target for metabolic disorders. Here, we present the molecular modeling, in vitro and in vivo characterization of Activator-3, 2-[2-(4-(trifluoromethyl)phenylamino)thiazol-4-yl]acetic acid, an AMP mimetic and a potent pan-AMPK activator. Activator-3 and AMP likely share common activation mode for AMPK activation. Activator-3 enhanced AMPK phosphorylation by upstream kinase LKB1 and protected AMPK complex against dephosphorylation by PP2C. Molecular modeling analyses followed by in vitro mutant AMPK enzyme assays demonstrate that Activator-3 interacts with R70 and R152 of the CBS1 domain on AMPK γ subunit near AMP binding site. Activator-3 and C2, a recently described AMPK mimetic, bind differently in the γ subunit of AMPK. Activator-3 unlike C2 does not show cooperativity of AMPK activity in the presence of physiological concentration of ATP (2 mM). Activator-3 displays good pharmacokinetic profile in rat blood plasma with minimal brain penetration property. Oral treatment of High Sucrose Diet (HSD) fed diabetic rats with 10 mg/kg dose of Activator-3 once in a day for 30 days significantly enhanced glucose utilization, improved lipid profiles and reduced body weight, demonstrating that Activator-3 is a potent AMPK activator that can alleviate the negative metabolic impact of high sucrose diet in rat model.

The effects of naringenin and naringin on the glucose uptake and AMPK phosphorylation in high glucose treated HepG2 cells

“…The phosphorylation of Thr172 within the activation loop of the α-subunit can cause AMPK activation [ 7 ]. AMP is a natural activator of AMPK and binds to the CBS domains of the γ-subunit and indirectly promotes the activity of the catalytic domain in α-subunit [ 23 ]. The binding of AMP to the γ-subunit of AMPK stimulates the phosphorylation of Thr172 in the α-subunit by upstream kinases LKB1 and CaMKKβ, and prevents the dephosphorylation of Thr172 in α-subunit [ 24 25 ].…”

Evaluation of Biosafety, Antiobesity, and Endothelial Cells Proliferation Potential of Basil Seed Extract Loaded Organic Solid Lipid Nanoparticle

“…The lipolytic effect of BSE-SLNp might be due to the bioactive principle, 2,4-dinitro-N2,N3-dipropyl-6-(trifluoromethyl)-1,3-benzenediamine and gibberellic acid present in BSE, which is the stimulator of the β-adrenergic receptor followed by the activation of AMPK, effectively increasing the protein expression levels of CREBp-1 and AMPK. In this context, Bung et al (2018) have established that 3, 2-[2-(4-(trifluoromethyl)phenylamino) thiazol-4-yl] acetic acid was effective as an AMP mimetic or AMPK activator in vitro and in vivo . cAMP pathways and AMP-activated protein kinase (AMPK) were recognized as cellular energy sensors vital for the reserve of adipocyte maturation ( Zhang et al, 2015 ).…”

AMP-activated protein kinase: the current landscape for drug development

“…Intriguingly, these binding sites differ from the nucleotide binding sites, suggesting that the γ subunit could be exploited for drug targeting independently of nucleotide binding 170 . In this regard, a recent study identified 2-(2-(4-(trifluoromethyl)phenylamino)thiazol-4-yl)acetic acid as a potent AMPK activator that acts as an AMP mimetic 171 . Another small molecule, O304, has recently been shown to protect against AMPK phosphorylated Thr172 (pThr172) dephosphorylation by protein phosphatase 2C without allosteric activation of AMPK, thus mimicking the effects of ADP but not AMP 28 .…”