Using the standard patch clamp whole cell recording method, we assessed the pharmacological activity of four fenamate nonsteroidal anti-inflammatory drugs, meclofenamic acid, flufenamic acid, mefenamic acid and niflumic acid, on hKv2.1, a major human neuronal voltage-gated potassium channel stably expressed heterologously in Chinese hamster ovary cells. Meclofenamic acid inhibited hKv2.1 in a concentration-dependent manner whereas the other three fenamates had weaker or no effect on these channels at a concentration of 100 microM. The estimated IC50 of meclofenamic acid was 56.0 microM for hKv2.1 compared an IC50 of 155.9 microM for another human neuronal K channel (hKv1.1). Meclofenamic acid reached its maximum inhibition within 5 min of bath application and its effect was readily reversed upon wash. Kinetic analysis revealed that this drug did not alter the channel activation or deactivation time courses. Moreover, the effect of meclofenamic acid on hKv2.1 channels was not voltage-dependent. Indomethacin, another inhibitor of the cyclooxygenase that catalyses the synthesis of prostaglandin from arachidonic acid, had no effect on either hKv2.1 or hKv1.1. These results indicate that meclofenamic acid inhibits hKv2.1 more potently than hKv1.1 and it is likely that this compound acts directly on the channel proteins.