Identification of DGAT2 Inhibitors Using Mass Spectrometry

J Biomol Screen. 2016 Feb;21(2):117-26. doi: 10.1177/1087057115607463. Epub 2015 Sep 24.


Mass spectrometry offers significant advantages over other detection technologies in the areas of hit finding, hit validation, and medicinal chemistry compound optimization. The foremost obvious advantage is the ability to directly measure enzymatic product formation. In addition, the inherent sensitivity of the liquid chromatography/mass spectrometry (LC/MS) approach allows the execution of enzymatic assays at substrate concentrations typically at or below substrate Km. Another advantage of the LC/MS approach is the ability to assay impure enzyme systems that would otherwise be difficult to prosecute with traditional labeled methods. This approach was used to identify inhibitors of diacylglycerol O-acyltransferase-2 (DGAT2), a transmembrane enzyme involved in the triglyceride (TG) production pathway. The LC/MS approach was employed because of its increased assay window (compared with control membranes) of more than sevenfold compared with less than twofold with a conventional fluorogenic assay. The ability to generate thousands of dose-dependent IC50 data was made possible by the use of a staggered parallel LC/MS system with fast elution gradients. From the hit-deconvolution efforts, several structural scaffold series were identified that inhibit DGAT2 activity. Additional profiling of one chemotype in particular identified two promising reversible and selective compounds (compound 15 and compound 16) that effectively inhibit TG production in mouse primary hepatocytes.

Keywords: DGAT2; LC/MS; triglycerides.

MeSH terms

  • Animals
  • Cell Line
  • Chromatography, Liquid / methods
  • Diacylglycerol O-Acyltransferase / antagonists & inhibitors*
  • Diacylglycerol O-Acyltransferase / chemistry
  • Enzyme Assays / methods
  • Enzyme Inhibitors / chemistry*
  • Humans
  • Mass Spectrometry / methods
  • Sf9 Cells
  • Triglycerides / chemistry


  • Enzyme Inhibitors
  • Triglycerides
  • DGAT2 protein, human
  • Diacylglycerol O-Acyltransferase