Different Methods for Testing Potential Cyclooxygenase-1 and Cyclooxygenase-2 Inhibitors

Abstract

The need for the development of selective agents, which only inhibit the mainly “harmful” cyclooxygenase-2 (COX-2) while leaving physiological COX-1 mostly unaffected, still remains, especially after the recent issues related to cardiovascular toxicity caused by some COX-2 selective agents. Thus there is still a demand for sensitive and rapid methods to assay for COX-2 selective agents. Among several in vitro testing systems the whole blood assay (WBA) is a well-known method to examine non-steroidal anti-inflammatory drugs (NSAIDs) in view of their potency to inhibit COX activity. This assay has some major advantages over enzyme-based or isolated cell assays. Emergence of artifacts due to cell separation steps is kept to a minimum and substances, even in disproportional high concentrations, can be examined outside the body in a physiological environment resembling most closely the in vivo conditions in living humans, i.e., 37°C, homeostasis, presence of all blood compounds and cell–cell interactions remain intact. While COX-1 human whole blood assays are performed within less than 2 h, for established COX-2 assays one still has to allow for an overnight incubation step before gaining the desired plasma. The aim of the assay described in this chapter is to characterize an optimized human whole blood assay (hWBA). We present a simple, fast and reliable method to examine the capacity of NSAIDs at inhibiting COX-2 activity that can be applied for rapid and routine screening purposes.