Modification of the Orthosteric PPAR? Covalent Antagonist Scaffold Yields an Improved Dual-Site Allosteric Inhibitor.

ABSTRACT: GW9662 and T0070907 are widely used commercially available irreversible antagonists of peroxisome proliferator-activated receptor gamma (PPAR?). These antagonists covalently modify Cys285 located in an orthosteric ligand-binding pocket embedded in the PPAR? ligand-binding domain and are used to block binding of other ligands. However, we recently identified an alternate/allosteric ligand-binding site in the PPAR? LBD to which ligand binding is not inhibited by these orthosteric covalent antagonists. Here, we developed a series of analogs based on the orthosteric covalent antagonist scaffold with the goal of inhibiting both orthosteric and allosteric cellular activation of PPAR? by MRL20, an orthosteric agonist that also binds to an allosteric site. Our efforts resulted in the identification of SR16832 (compound 22), which functions as a dual-site covalent inhibitor of PPAR? transcription by PPAR?-binding ligands. Molecular modeling, protein NMR spectroscopy structural analysis, and biochemical assays indicate the inhibition of allosteric activation occurs in part through expansion of the 2-chloro-5-nitrobenzamidyl orthosteric covalent antagonist toward the allosteric site, weakening of allosteric ligand binding affinity, and inducing conformational changes not competent for cellular PPAR? activation. Furthermore, SR16832 better inhibits binding of rosiglitazone, a thiazolidinedione (TZD) that weakly activates PPAR? when cotreated with orthosteric covalent antagonists, and may better inhibit binding of endogenous PPAR? ligands such as docosahexaenoic acid (DHA) compared to orthosteric covalent antagonists. Compounds such as SR16832 may be useful chemical tools to use as a dual-site bitopic orthosteric and allosteric covalent inhibitor of ligand binding to PPAR?.

SUBMITTER: Brust R 

PROVIDER: S-EPMC5652320 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature