Block of sensory neuronal Na+ channels by the secreolytic ambroxol is associated with an interaction with local anesthetic binding sites.

Voltage-gated Na(+) channels (Na(v)) regulate the excitability of sensory neurons and are potential targets for novel analgesics. The secreolytic ambroxol reduces pain-related behavior in rodents and alleviates pain in humans. With properties resembling those of local anesthetics, ambroxol has been reported to block Na(+) currents in sensory neurons with a preference for tetrodotoxin-resistant (TTXr) Na(+) currents encoded by Na(v)1.8. However, the molecular determinants for ambroxol-induced block of Na(+) channels and a preferential block of Na(v)1.8 opposed to tetrodotoxin-sensitive (TTXs) Na(v) alpha-subunits have not been studied in detail. By means of whole-cell voltage clamp recordings, we studied the effects of ambroxol and local anesthetics on the recombinant TTXr subunit Na(v)1.8, on TTXs Na(v) alpha-subunits and on mutants of Na(v)1.4 that are insensitive to local anesthetics. Tonic and use-dependent block by ambroxol was strongly alleviated in local anesthetic-insensitive Na(v)1.4 mutants. Use-dependent block, but not tonic block was significantly stronger on Na(v)1.8 than on TTXs channels. The TTXs subunit Na(v)1.3 displayed the least degree of use-dependent block by ambroxol. The local anesthetics mepivacaine and S(-)-bupivacaine also blocked Na(v)1.8 and TTXs channels differentially. While mepivacaine displayed a preferential use-dependent block of Na(v)1.8, S(-)-bupivacaine displayed a preference for TTXs Na(+) channels. Our data show that ambroxol acts as a typical local anesthetic on Na(+) channels interacting with specific residues in the S6 segments. This property probably meditates the analgesic effect of ambroxol. Ambroxol preferentially blocks Na(v)1.8, however shares this property with established local anesthetics like mepivacaine.