Discovery of Human Intestinal MGAT Inhibitors Using High-Throughput Mass Spectrometry

SLAS Discov. 2017 Apr;22(4):360-365. doi: 10.1177/1087057116673181. Epub 2016 Oct 8.


Monoacylglycerol acyltransferase (MGAT) activity catalyzes the synthesis of diacylglycerol (DAG) from fatty acyl-CoA and monoacylglycerol as substrates. It is important for the resynthesis of triacylglycerol (TAG) in the intestine. In the present study, we developed a MGAT enzymatic assay of human intestinal microsomes using a high-throughput mass spectrometry (MS)-based detection system. After screening with small-molecular-weight libraries for compounds exhibiting inhibitions against DAG and the consequent TAG syntheses, we identified multiple compounds that specifically inhibit intestinal MGAT activity. The inhibitory activities of these compounds were correlated to those determined using a recombinant human MGAT2 enzyme. An aryl-sulfonamide compound T1 showed potent inhibitory activity toward human intestinal MGAT and recombinant human MGAT2, with selectivity over MGAT3. This high-throughput MS-based assay provides a novel platform for the discovery of DAG or TAG synthesis inhibitors. The identified aryl-sulfonamide compound T1 is a promising starting compound for optimization studies of inhibitors with selectivity toward MGAT2.

Keywords: enzyme assays or enzyme kinetics; lipids or lipid; mass spectrometry; metabolism; multiplex assays and technology.

MeSH terms

  • Acyl Coenzyme A / metabolism
  • Animals
  • COS Cells
  • Chlorocebus aethiops
  • Diglycerides / antagonists & inhibitors*
  • Diglycerides / biosynthesis
  • Drug Discovery
  • Enzyme Inhibitors / chemistry
  • Enzyme Inhibitors / pharmacology*
  • Gene Expression
  • High-Throughput Screening Assays*
  • Humans
  • Intestines / drug effects
  • Intestines / enzymology
  • Mass Spectrometry / methods
  • Microsomes / drug effects*
  • Microsomes / enzymology
  • Monoglycerides / metabolism
  • N-Acetylglucosaminyltransferases / antagonists & inhibitors
  • N-Acetylglucosaminyltransferases / genetics
  • N-Acetylglucosaminyltransferases / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Substrate Specificity
  • Sulfonamides / chemistry
  • Sulfonamides / pharmacology*
  • Triglycerides / antagonists & inhibitors*
  • Triglycerides / biosynthesis


  • 1,2-diacylglycerol
  • Acyl Coenzyme A
  • Diglycerides
  • Enzyme Inhibitors
  • Monoglycerides
  • Recombinant Proteins
  • Sulfonamides
  • Triglycerides
  • N-Acetylglucosaminyltransferases
  • alpha-1,6-mannosyl-glycoprotein beta-1,2-N-acetylglucosaminyltransferase
  • beta-1,4-mannosyl-glycoprotein beta-1,4-N-acetylglucosaminyltransferase