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

Fowkes, Milan M., Troeberg, Linda ORCID:, Brennan, Paul E., Vincent, Tonia L., Meldal, Morten and Lim, Ngee H. (2023) Development of selective ADAMTS-5 peptide substrates to monitor proteinase activity. Journal of Medicinal Chemistry, 66 (5). pp. 3522-3539. ISSN 0022-2623

[thumbnail of acs.jmedchem.2c02090]
PDF (acs.jmedchem.2c02090) - Published Version
Available under License Creative Commons Attribution.

Download (8MB) | Preview


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.

Item Type: Article
Additional Information: Funding Information: This study was funded by the Kennedy Trust for Rheumatology Research through a Kennedy Trust Prize Studentship awarded to Dr. Fowkes to perform his DPhil at the University of Oxford. The combinatorial chemistry part of the project was carried out in the Department of Chemistry at the University of Copenhagen and was funded by a Researcher Mobility Grant awarded to Dr Fowkes by the Royal Society of Chemistry. Additional funding was provided by the Centre for OA Pathogenesis Versus Arthritis (grant numbers 21621 and 20205). Publisher Copyright: © 2023 The Authors. Published by American Chemical Society
Uncontrolled Keywords: molecular medicine,drug discovery ,/dk/atira/pure/subjectarea/asjc/1300/1313
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
UEA Research Groups: Faculty of Medicine and Health Sciences > Research Centres > Metabolic Health
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 14 Jun 2023 14:15
Last Modified: 19 Oct 2023 03:36
DOI: 10.1021/acs.jmedchem.2c02090


Downloads per month over past year

Actions (login required)

View Item View Item