Development of Selective ADAMTS-5 Peptide Substrates to Monitor Proteinase Activity

J Med Chem. 2023 Mar 9;66(5):3522-3539. doi: 10.1021/acs.jmedchem.2c02090. Epub 2023 Feb 22.


The dysregulation of proteinase activity is a hallmark of osteoarthritis (OA), a disease characterized by progressive degradation of articular cartilage by catabolic proteinases such as a disintegrin and metalloproteinase with thrombospondin type I motifs-5 (ADAMTS-5). The ability to detect such activity sensitively would aid disease diagnosis and the evaluation of targeted therapies. Förster resonance energy transfer (FRET) peptide substrates can detect and monitor disease-related proteinase activity. To date, FRET probes for detecting ADAMTS-5 activity are nonselective and relatively insensitive. We describe the development of rapidly cleaved and highly selective ADAMTS-5 FRET peptide substrates through in silico docking and combinatorial chemistry. The lead substrates 3 and 26 showed higher overall cleavage rates (∼3-4-fold) and catalytic efficiencies (∼1.5-2-fold) compared to the best current ADAMTS-5 substrate ortho-aminobenzoyl(Abz)-TESE↓SRGAIY-N-3-[2,4-dinitrophenyl]-l-2,3-diaminopropionyl(Dpa)-KK-NH2. They exhibited high selectivity for ADAMTS-5 over ADAMTS-4 (∼13-16-fold), MMP-2 (∼8-10-fold), and MMP-9 (∼548-2561-fold) and detected low nanomolar concentrations of ADAMTS-5.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAMTS4 Protein / metabolism
  • ADAMTS5 Protein / metabolism
  • Cartilage, Articular* / metabolism
  • Endopeptidases / metabolism
  • Humans
  • Osteoarthritis* / metabolism
  • Peptides / metabolism
  • Proteolysis


  • Peptides
  • Endopeptidases
  • ADAMTS4 Protein
  • ADAMTS5 Protein