harvested as described above and analyzed by elisa for levels of il 6 endogen eh2 il6 and il 8 endogen eh2 il8 according to the instructions of the manufacturer the elisa data are used to generate dose response curves from which ic50 values are derived e inhibition of lps induced prostaglandin endoperoxide synthase 2 pghs 2 or cox 2 induction in pbmcs human peripheral mononuclear cells pbmcs are isolated from fresh human blood buffy coats by centrifugation in a vacutainer cpt becton amp dickinson 15 u00d7 10 6 cells are seeded in a 6 well tissue culture dish containing rpmi 1640 supplemented with 10 fetal bovine serum 50 u ml penicillin 50 u03bcg ml streptomycin and 2 mm l glutamine an inhibitor of the instant invention is added at 0 2 2 0 and 20 u03bcm final concentrations in dmso then lps is added at a final concentration of 4 ng ml to induce enzyme expression the final culture volume is 10 ml well after overnight incubation at 37 u00b0 c 5 co2 the cells are harvested by scraping and subsequent centrifugation then the supernatant is removed and the cells are washed twice in ice cold dpbs dulbecco 39 s phosphate buffered saline.