Identification of DGAT2 Inhibitors Using Mass Spectrometry.

Autor:	Song XS; Department of Pharmacology, Merck Research Laboratories, Boston, MA, USA., Zhang J; Department of Pharmacology, Merck Research Laboratories, Kenilworth, NJ, USA., Chen X; Department of Pharmacology, Merck Research Laboratories, Kenilworth, NJ, USA., Palyha O; Atherosclerosis, Merck Research Laboratories, Kenilworth, NJ, USA., Chung C; Department of Pharmacology, Merck Research Laboratories, Kenilworth, NJ, USA., Sonatore LM; Department of Pharmacology, Merck Research Laboratories, Kenilworth, NJ, USA., Wilsie L; Atherosclerosis, Merck Research Laboratories, Kenilworth, NJ, USA., Stout S; Department of Pharmacology, Merck Research Laboratories, Kenilworth, NJ, USA., McLaren DG; Department of Pharmacology, Merck Research Laboratories, Kenilworth, NJ, USA., Taggart A; Atherosclerosis, Merck Research Laboratories, Kenilworth, NJ, USA., Imbriglio JE; Discovery Chemistry, Merck Research Laboratories, Kenilworth, NJ, USA., Pinto S; Atherosclerosis, Merck Research Laboratories, Kenilworth, NJ, USA., Garcia-Calvo M; Department of Pharmacology, Merck Research Laboratories, Kenilworth, NJ, USA., Addona GH; Department of Pharmacology, Merck Research Laboratories, Boston, MA, USA george_addona@merck.com.
Jazyk:	angličtina
Zdroj:	Journal of biomolecular screening [J Biomol Screen] 2016 Feb; Vol. 21 (2), pp. 117-26. Date of Electronic Publication: 2015 Sep 24.
DOI:	10.1177/1087057115607463
Abstrakt:	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. (© 2015 Society for Laboratory Automation and Screening.)
Databáze:	MEDLINE
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