Ecopladib

Male Sprague−Dawley rats are anesthetized, and 10−20 mL of filtered air is injected subcutaneously under the dorsal skin to form a pouch. Three and six days later, the pouches are reinflated with 10−15 mL of sterile air. On the seventh day, the test compound is dissolved in vehicle (55.5% Phosal 53 MCT, 5.6% Tween 80, 16.7% Labrasol, and 22.2% propylene carbonate) to give 37.5 mg of test compound per mL of vehicle. This test compound in vehicle is diluted with water to the appropriate concentration and dosed at 4 mL/kg. Vehicle treated animals receives the same amount of vehicle as the animals treated with the highest dose of compound. Two hours later, 2 mL of a 1% solution of carrageenan (Viscarin carrageenan type GP-209NF) in saline is injected into the pouch. Six hours after the carrageenan injection, the rats are individually sacrificed and the contents of the pouch are harvested. The amount of fluid recovered is measured. An aliquot of the exudate is centrifuged at 6500 rpm for 10 min, and 300 μL of each supernatant is precipitated with MeOH (800 μL) precooled to 0°C. The samples are well vortexed and are kept at −80°C overnight. The samples are centrifuged again and assayed for PGE2 to locate the PGE2 production within the linear range of the PGE2 standard curve. To minimize the difference in binding environments for the standards and samples, the standard curve is generated in a 1% solution of carrageenan that is mixed with assay buffer to the same dilution as the samples. The approximate ED₅₀ value is extrapolated from the dose−response curve.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.