Evaluation of a Putative PPAR Modulator

Abstract

BackgroundPeroxisome Proliferative‐Activated Receptors (PPAR) are part of a superfamily of receptors. Of the PPARs, three forms are most prevalent, PPAR‐a, PPAR‐b/d and PPAR‐g that bind FA & FA‐derivatives to their receptor domains. Each PPAR consists of an AF‐1 domain, DNA‐binding domain, dimerization domain, and ligand‐binding domain. (1) When a lipid‐derived ligand binds to a PPAR, the PPAR heterodimerizes with a retinoid X receptor (2). This heterodimer will bind a co‐activator protein (3), which in turn activates a peroxisome proliferator response element (PPRE) (1,2). The PPRE is located in a promoter region of the target genes of a DNA sequence that will allow for trans‐activation or ‐repression of gene expression. (4.) Previous data has been collected regarding a drug compound called Compound 9, which was a dual agonist of PPARg and PPARd. The compound tested, benzyl derivative, called BMZ is a derivative of compound 9.ObjectivesTest novel compound, a demethylated derivative of a previously used compound, called BMZ, to determine its activation of PPAR receptors.MethodsTo test BMZ activation of PPAR receptors, HeLa cells were plated into 96 well plates and transfected with PPRE‐3x‐TK‐luciferase (520ng/l), Trans IT‐X2 transfection reagent, and serum reduced media. The cells were then treated with troglitazone, pioglitazone, and BMZ with the following concentration in separate wells in triplicate, 0.1mM, 0.3 mM, 1.0 mM, 3.0 mM, 10 mM, 30 mM. Triplicate wells were also treated with 10 mM of BMZ with 10 mM of troglitazone. Finally, a separate triplicate of Compound 9 was added. A luciferase assay was conducted 24 hours after application of drug in five, five minute intervals.ResultsThe results showed that there was some activation of PPAR receptors for the novel BMZ compound. Furthermore, when BMZ was added to troglitazone, there was an decreased activation of the receptors.Support or Funding InformationMarian University College of Osteopathic Medicine Internal GrantRelative Luminescence Units of various PPAR‐gamma agonistsFigure 1