TY - JOUR

T1 - Development of Selective ADAMTS-5 Peptide Substrates to Monitor Proteinase Activity

AU - Fowkes, Milan M.

AU - Troeberg, Linda

AU - Brennan, Paul E.

AU - Vincent, Tonia L.

AU - Meldal, Morten

AU - Lim, Ngee H.

N1 - 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

PY - 2023

Y1 - 2023

N2 - 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.

AB - 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.

U2 - 10.1021/acs.jmedchem.2c02090

DO - 10.1021/acs.jmedchem.2c02090

M3 - Journal article

C2 - 36891740

AN - SCOPUS:85148874850

VL - 66

SP - 3522

EP - 3539

JO - Journal of Medicinal Chemistry

JF - Journal of Medicinal Chemistry

SN - 0022-2623

IS - 5

ER -