Human whole blood assay for rapid and routine testing of non-steroidal anti-inflammatory drugs (NSAIDs) on cyclo-oxygenase-2 activity

Inflammopharmacology. 2008 Aug;16(4):155-61. doi: 10.1007/s10787-008-8007-x.

Abstract

Aim of this study was to present a simple, fast and reliable method to examine the capacity of NSAIDs to inhibiting COX-2 activity that uses rapid (stimulation takes only 5 h compared to other existing protocols) and routine testing. The assay includes elimination of COX-1-activity using ASS (a selective COX-1 inhibitor) and the thromboxane synthetase inhibitor (TXBSI), COX-2 induction via LPS and measurement of PGE(2). Using TXBSI reduces the amount of LPS and results in higher prostaglandin production. Cremophor EL-EtOH was used as vehicle instead of DMSO because within a defined concentration range, Cremophor EL-EtOH allows even very hydrophobic drugs to be solubilized and applied in vitro without cell damage. Cremophor EL-EtOH at 0.2% was optimal as at this relatively low concentration excellent drug dissolution was obtained whereas many hydrophobic substances precipitate in 0.2% DMSO. Our results demonstrate that the IC(50) values for the tested NSAIDs are in the range of published data.

MeSH terms

  • Anti-Inflammatory Agents, Non-Steroidal / administration & dosage
  • Anti-Inflammatory Agents, Non-Steroidal / pharmacology*
  • Biological Assay / methods
  • Cyclooxygenase 2 / drug effects*
  • Cyclooxygenase 2 / metabolism
  • Cyclooxygenase 2 Inhibitors / administration & dosage
  • Cyclooxygenase 2 Inhibitors / pharmacology*
  • Ethanol / chemistry
  • Glycerol / analogs & derivatives
  • Glycerol / chemistry
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Inhibitory Concentration 50
  • Lipopolysaccharides / metabolism
  • Pharmaceutical Vehicles / chemistry
  • Reproducibility of Results
  • Solubility

Substances

  • Anti-Inflammatory Agents, Non-Steroidal
  • Cyclooxygenase 2 Inhibitors
  • Lipopolysaccharides
  • Pharmaceutical Vehicles
  • Ethanol
  • cremophor EL
  • Cyclooxygenase 2
  • Glycerol