BACKGROUND: Arachidyl amido cholanoic acid (Aramchol) is a potent downregulator 
of hepatic stearoyl-CoA desaturase 1 (SCD1) protein expression that reduces 
liver triglycerides and fibrosis in animal models of steatohepatitis. In a phase 
IIb clinical trial in patients with nonalcoholic steatohepatitis (NASH), 52 wk 
of treatment with Aramchol reduced blood levels of glycated hemoglobin A1c, an 
indicator of glycemic control.
AIM: To assess lipid and glucose metabolism in mouse hepatocytes and in a NASH 
mouse model [induced with a 0.1% methionine and choline deficient diet (0.1MCD)] 
after treatment with Aramchol.
METHODS: Isolated primary mouse hepatocytes were incubated with 20 μmol/L 
Aramchol or vehicle for 48 h. Subsequently, analyses were performed including 
Western blot, proteomics by mass spectrometry, and fluxomic analysis with 
13C-uniformly labeled glucose. For the in vivo part of the study, male C57BL/6J 
mice were randomly fed a control or 0.1MCD for 4 wk and received 1 or 5 mg/kg/d 
Aramchol or vehicle by intragastric gavage for the last 2 wk. Liver metabolomics 
were assessed using ultra-high-performance liquid chromatography-time of 
flight-MS for the determination of glucose metabolism-related metabolites.
RESULTS: Combination of proteomics and Western blot analyses showed increased 
AMPK activity while the activity of nutrient sensor mTORC1 was decreased by 
Aramchol in hepatocytes. This translated into changes in the content of their 
downstream targets including proteins involved in fatty acid (FA) synthesis and 
oxidation [P-ACCα/β(S79), SCD1, CPT1A/B, HADHA, and HADHB], oxidative 
phosphorylation (NDUFA9, NDUFB11, NDUFS1, NDUFV1, ETFDH, and UQCRC2), 
tricarboxylic acid (TCA) cycle (MDH2, SUCLA2, and SUCLG2), and ribosome 
(P-p70S6K[T389] and P-S6[S235/S236]). Flux experiments with 13C-uniformely 
labeled glucose showed that TCA cycle cataplerosis was reduced by Aramchol in 
hepatocytes, as indicated by the increase in the number of rounds that malate 
remained in the TCA cycle. Finally, liver metabolomic analysis showed that 
glucose homeostasis was improved by Aramchol in 0.1MCD fed mice in a 
dose-dependent manner, showing normalization of glucose, G6P, F6P, UDP-glucose, 
and Rbl5P/Xyl5P.
CONCLUSION: Aramchol exerts its effect on glucose and lipid metabolism in NASH 
through activation of AMPK and inhibition of mTORC1, which in turn activate FA 
β-oxidation and oxidative phosphorylation.