Kinase experiment:

A homogeneous, fluorescent polarization PPARα and PPARγ binding assay is used as the primary screen for determining the PPARα and PPARγ binding affinity of compounds. The human functional activity of PPARα and PPARγ agonists is determined by using the GAL4-LBD assays. The in vitro hamster, rat, and mouse PPARα functional activities are tested in the chimeric GAL4/PPARα assay format. The data are reported as an EC50 value calculated using XLfit 4 parameter fit and floating all parameters. Full length human PPARα and PPARγ co-transfection assays in HepG2 cells are employed for further testing the leading compounds (BMS-687453)[1].

Animal experiment:

Male 6–8 week old human apoA1 transgenic mice are randomly assigned into different treatment groups and weighed and dosed by oral gavage (5 mL/kg body weight) once a day in the morning with vehicle alone or with compound (BMS-687453) and allowed free access to food and water. The study duration is 10 days. After dosing on day 10, mice are fasted for 4 h and sacrificed by CO2 asphyxiation, and blood samples are collected in serum-separating tubes via cardiac puncture for lipid measurements. Livers are dissected out, weighed, and quickly frozen in liquid nitrogen for future RNA analysis. Human apoA1 concentration in serum is measured using the apolipoprotein A1 kit[1].

IC50: 10 nM for human PPARα

BMS 687453 is a PPARα agonist.

The peroxisome proliferator activated receptor (PPAR) is a member of the intracellular nuclear hormone receptor superfamily of transcription factors, having pleiotropic effects on plasma lipoprotein levels, insulin sensitization, atherosclerosis, and inflammation.

In vitro: BMS-687453 was identified as a potent and selective PPAR alpha agonist, with approximately 410-fold selectivity versus human PPARgamma in PPAR-GAL4 transactivation assays. In addition, similar potency and selectivity were also seen in the full length receptor co-transfection assays. [1].

In vivo: In previous study, BMS-687453 had an excellent pharmacokinetic profile across all tested animal species. The oral absorption was rapid in mouse, rat, dog, and cynomulgus monkey. BMS-687453 also exhibited low plasma clearance in the mouse, rat, and monkey and moderate plasma clearance in the dog. The half-life of BMS-687453 ranged from 3 h in mouse to 12 h in cynomolgus monkeys. BMS-687453 showed excellent absolute oral bioavailability ranging from 58% (dog) to 91% (rat) [1].

Clinical trial: Up to now, BMS 687453 is still in the preclinical development stage.

Reference:
[1] Li J.  Discovery of an oxybenzylglycine based peroxisome proliferator activated receptor alpha selective agonist 2-((3-((2-(4-chlorophenyl)-5-methyloxazol-4-yl)methoxy)benzyl)(methoxycarbonyl) amino)acetic acid (BMS-687453). J Med Chem. 2010 Apr 8;53(7):2854-64.