Mechanisms of acid-sensing ion channels inhibition by nafamostat, sepimostat and diminazene

Acid-sensing ion channels (ASICs) are blocked by many cationic compounds. Mechanisms of action, which may include pore block, modulation of activation and desensitization, need systematic analysis to allow predictable design of new potent and selective drugs. In this work, we studied the action of the serine protease inhibitors nafamostat, sepimostat, gabexate and camostat, on native ASICs in rat giant striatal interneurons and recom-binant ASIC1a and ASIC2a channels, and compared it to that of well-known small molecule ASIC blocker diminazene. All these compounds have positively charged amidine and/or guanidine groups in their structure. Nafamostat, sepimostat and diminazene inhibited pH 6.5-induced currents in rat striatal interneurons at-80 mV holding voltage with IC50 values of 0.78 +/- 0.12 mu M, 2.4 +/- 0.3 mu M and 0.40 +/- 0.09 mu M, respectively, whereas camostat and gabexate were practically ineffective. The inhibition by nafamostat, sepimostat and diminazene was voltage-dependent evidencing binding in the channel pore. They were not trapped in the closed channels, suggesting foot-in-the-door mechanism of action. The inhibitory activity of nafamostat, sepimostat and diminazene was similar in experiments on native ASICs and recombinant ASIC1a channels, while all of them were drastically less active against ASIC2a channels. According to our molecular modeling, three active com-pounds bind in the channel pore between Glu 433 and Ala 444 in a similar way. In view of the relative safety of nafamostat for clinical use in humans, it can be considered as a potential candidate for the treatment of path-ophysiological conditions linked to ASICs disfunction, including inflammatory pain and ischemic stroke.

Automated summary

This study investigated the mechanisms of action of various cationic compounds on acid-sensing ion channels (ASICs) and proposed a predictive analysis method for designing new drugs. Among them, nafamostat showed potential application value in the treatment of diseases related to ASIC dysfunction.