N-(p-amylcinnamoyl) Anthranilic Acid (ACA) is a broad spectrum Phospholipase A2 (PLA2) inhibitor and TRP channel blocker.

ACA modulates different TRP channels most probably by a direct interaction. Owing to its high potency and efficacy on TRPM2, ACA can serve, in combination with other blockers, as a pharmacological tool for studying H2O2-induced Ca2+ signalling and biological functions of TRPM2 channels in native cells[1].ACA is also an inhibitor of phospholipase A2, blocking the release of arachidonic acid when given at 50 μM[2].

Measurements of [Ca2+]i were carried out on TRPM8-transfected HEK293 cells grown in 96-well microplates. Cells were loaded with 1?μm fluo-4 for 30?min at 37°C in a standard solution. Subsequently, the standard solution was exchanged for a solution containing 1?mm EGTA instead of 2?mm CaCl2, and plates were inserted into a fluorometric-imaging plate reader. The fluorescence was excited at 485?nm, emitted at 535?nm and corrected for the background fluorescence. Fluorescence values were measured after addition of ACA, menthol and 3?mm CaCl2 to the EGTA-containing solution. ACA, menthol and 3?mm CaCl2 were applied consecutively in the given order with a time delay of 2?min between each application[1].