Development of selective ADAMTS-5 peptide substrates to monitor proteinase activity

Journal article

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.

Authors: Fowkes, MM; Troeberg, L; Brennan, PE; Vincent, TL; Meldal, M

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: American Chemical Society
• Journal: Journal of Medicinal Chemistry More from this journal
• Volume: 66
• Issue: 5
• Pages: 3522-3539
• Publication date: 2023-02-22
• Acceptance date: 2023-02-22
• DOI: 10.1021/acs.jmedchem.2c02090
• EISSN: 1520-4804
• ISSN: 0022-2623
• Pmid: 36891740
• Language: English
• Keywords: articular cartilage; proteolysis; ADAMTS4 protein; osteoarthritis; peptides; endopeptidases; FFR