Pharmacological activity and NMR solution structure of the leech peptide HSTX-I

The role of voltage-gated sodium (Na_V) channels in pain perception is indisputable. Of particular interest as targets for the development of pain therapeutics are the tetrodotoxin-resistant isoforms Na_V1.8 and Na_V1.9, based on animal as well as human genetic studies linking these ion channel subtypes to the pathogenesis of pain. However, only a limited number of inhibitors selectively targeting these channels have been reported. HSTX-I is a peptide toxin identified from saliva of the leech Haemadipsa sylvestris. The native 23-residue peptide, stabilised by two disulfide bonds, has been reported to inhibit rat Na_V1.8 and mouse Na_V1.9 with low micromolar activity, and may therefore represent a scaffold for development of novel modulators with activity at human tetrodotoxin-resistant Na_V isoforms. We synthetically produced this hydrophobic peptide in high yield using a one-pot oxidation and single-step purification and determined the three-dimensional solution structure of HSTX-I using NMR solution spectroscopy. However, in our hands, the synthetic HSTX-I displayed only very modest activity at human Na_V1.8 and Na_V1.9 and lacked analgesic efficacy in a murine model of inflammatory pain.