Differential inhibition of murine prostaglandin synthase-1 and -2 by nonsteroidal anti-inflammatory drugs using exogenous and endogenous sources of arachidonic acid.
Mouse embryonic fibroblasts (10T1/2) and Chinese hamster ovary (AS52) cell lines that stably express murine prostaglandin G/H synthase (PGHS)-1 or -2 were used to compare the effects of exogenous and endogenous arachidonic acid (AA) on isozyme-selective inhibition by acetylsalicylic acid, indomethacin, and N-[2-cyclohexyloxyl-4-nitrophenyl] methanesulfonamide (NS-398). The rationale for developing in vitro systems that identify PGHS-2-selective inhibitors is the belief that inhibition of this isoform accounts for the therapeutic benefits of nonsteroidal anti-inflammatory drugs (NSAIDs). Conversely, inhibition of PGHS-1 is believed to cause the toxic effects of NSAIDs, such as gastric and renal damage. When exogenous AA was used, acetylsalicylic acid was a 5- to 10-fold more potent inhibitor of PGHS-1, whereas indomethacin was a 4- to 5-fold more potent inhibitor of PGHS-2. Within the dose range tested (1 x 10(-6) microM to 100 microM), NS-398 was highly selective for PGHS-2. When calcium ionophore A23187 was used to mobilize endogenous AA, acetylsalicylic acid and indomethacin equipotently inhibited both PGHS-1 and PGHS-2 isozymes. NS-398 remained highly selective for PGHS-2 in 10T1/2 and AS52 cells but also effectively (100%) inhibited PGHS-1 in AS52 cells. Pharmacological data derived using endogenous AA correlated better with the anti-inflammatory efficacy of these NSAIDs in laboratory animals and with the therapeutic/toxic activities of these NSAIDs in rheumatoid arthritic patients. Therefore, screening for PGHS-selective NSAIDs may best be conducted in intact cells that express high levels of each isozyme using endogenous sources of AA.
Chulada
P C
PC
National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709, USA.
Langenbach
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R
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D016718:Arachidonic Acid / Q000378:metabolism*; D001241:Aspirin / Q000494:pharmacology; D016466:CHO Cells; D000001:Calcimycin / Q000494:pharmacology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D052246:Cyclooxygenase 2 Inhibitors; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology*; D007213:Indomethacin / Q000494:pharmacology; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins; D051379:Mice; D009578:Nitrobenzenes / Q000494:pharmacology; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism*; D013449:Sulfonamides / Q000494:pharmacology
D000894:Anti-Inflammatory Agents, Non-Steroidal; D052246:Cyclooxygenase 2 Inhibitors; D016861:Cyclooxygenase Inhibitors; D007527:Isoenzymes; D008565:Membrane Proteins; D009578:Nitrobenzenes; D013449:Sulfonamides; C080955:N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; D016718:Arachidonic Acid; D000001:Calcimycin; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; C496512:Ptgs1 protein, mouse; D001241:Aspirin; D007213:Indomethacin