Non-canonical amino acid scanning refines a flea-leech chimeric thrombin inhibitor
- A rapid, high-throughput workflow was set up to assemble many modified peptide analogues in parallel.
- Variants derive from a chimeric sulfopeptide combining a flea-derived fragment with a leech hirudin segment.
- Non-canonical residues were placed at selected positions to probe non-native structural motifs.
- Each analogue was assessed for thrombin inhibition and resistance to cleavage, linking sequence changes to activity.
💡 Points Based on the abstract
The sulfotyrosine is the liability to watch: in hirudin-type exosite-I binders the sulfated Tyr boosts affinity, yet sulfate esters are acid-labile and awkward to install in parallel. I'd want a head-to-head against bivalirudin and a plasma stability readout, not just buffer protease assays. Next step: swap in a sulfotyrosine mimetic (sulfonate or difluoromethylene phosphonate) to decouple potency from the labile ester.