title: Characterization of monoacylglycerol acyltransferase 2 inhibitors by a novel probe in binding assays.
abstract: Monoacylglycerol acyltransferase 2 (MGAT2) is a membrane-bound lipid acyltransferase that catalyzes the formation of diacylglycerol using monoacylglycerol and fatty acyl CoA as substrates. MGAT2 is important for intestinal lipid absorption and is an emerging target for the treatment of metabolic diseases. In the current study, we identified and characterized four classes of novel MGAT2 inhibitors. We established both steady state and kinetic binding assay protocols using a novel radioligand, [(3)H]compound A. Diverse chemotypes of MGAT2 inhibitors were found to compete binding of [(3)H]compound A to MGAT2, indicating the broad utility of [(3)H]compound A for testing various classes of MGAT2 inhibitors. In the dynamic binding assays, the kinetic values of MGAT2 inhibitors such as Kon, Koff, and T1/2 were systematically defined. Of particular value, the residence times of inhibitors on MGAT2 enzyme were derived. We believe that the identification of novel classes of MGAT2 inhibitors and the detailed kinetic characterization provide valuable information for the identification of superior candidates for in vivo animal and clinical studies. The current work using a chemical probe to define inhibitory kinetics can be broadly applied to other membrane-bound acyltransferases.
authors.affiliation: Department of Fibrosis Discovery, Research and Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Zhengping
authors.initials: Z
authors.lastname: Ma
authors.affiliation: Department of Discovery Chemistry, Research and Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Hannguang J
authors.initials: HJ
authors.lastname: Chao
authors.affiliation: Department of Discovery Chemistry, Research and Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Huji
authors.initials: H
authors.lastname: Turdi
authors.affiliation: Department of Discovery Chemistry, Research and Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Jon J
authors.initials: JJ
authors.lastname: Hangeland
authors.affiliation: Department of Discovery Chemistry, Research and Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Todd
authors.initials: T
authors.lastname: Friends
authors.affiliation: Department of Mechanistic Biochemistry, Research and Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Kopcho
authors.affiliation: Department of Discovery Chemistry, Research and Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: R Michael
authors.initials: RM
authors.lastname: Lawrence
authors.affiliation: Department of Fibrosis Discovery, Research and Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA. Electronic address: dong.cheng@bms.com.
authors.forename: Dong
authors.initials: D
authors.lastname: Cheng
mesh_terms: D000217:Acyltransferases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D000818:Animals; D004353:Drug Evaluation, Preclinical / Q000379:methods; D057075:Enzyme Assays / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D008024:Ligands; D051379:Mice; D011485:Protein Binding / Q000187:drug effects; D011869:Radioligand Assay / Q000379:methods; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D008024:Ligands; D011994:Recombinant Proteins; D000217:Acyltransferases; C021802:2-acylglycerol O-acyltransferase
keywords: Acyltransferase; Enzyme inhibitor binding assay; Enzyme inhibitor kinetics; MGAT2; Triglyceride synthesis