Centre for OA Pathogenesis Versus Arthritis, Kennedy Institute of Rheumatology, University of Oxford, Roosevelt Drive, Headington, Oxford OX3 7FY, United Kingdom.
Norwich Medical School, Bob Champion Research and Education Building, Rosalind Franklin Road, University of East Anglia, Norwich NR4 7UQ, United Kingdom.
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 docking and combinatorial chemistry. The lead substrates and showed higher overall cleavage rates (∼3-4-fold) and catalytic efficiencies (∼1.5-2-fold) compared to the best current ADAMTS-5 substrate -aminobenzoyl(Abz)-TESE↓SRGAIY--3-[2,4-dinitrophenyl]-l-2,3-diaminopropionyl(Dpa)-KK-NH. 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.
蛋白酶活性失调是骨关节炎 (OA) 的一个标志，OA 是一种疾病，其特征是通过分解代谢蛋白酶（如解整合素和金属蛋白酶与凝血酶反应素-1 型基序 5 [ADAMTS-5]）渐进性降解关节软骨。能够灵敏地检测到这种活性将有助于疾病诊断和靶向治疗的评估。Förster 共振能量转移 (FRET) 肽底物可用于检测和监测与疾病相关的蛋白酶活性。迄今为止，用于检测 ADAMTS-5 活性的 FRET 探针是非选择性的，相对不敏感。我们通过对接和组合化学描述了快速切割和高度选择性 ADAMTS-5 FRET 肽底物的开发。先导底物 和 与目前最好的 ADAMTS-5 底物 -aminobenzoyl(Abz)-TESE↓SRGAIY--3-[2,4-dinitrophenyl]-l-2,3-diaminopropionyl(Dpa)-KK-NH 相比，表现出更高的总切割速率（∼3-4 倍）和催化效率（∼1.5-2 倍）。它们对 ADAMTS-5 相对于 ADAMTS-4（∼13-16 倍）、MMP-2（∼8-10 倍）和 MMP-9（∼548-2561 倍）具有高选择性，并检测到低纳摩尔浓度的 ADAMTS-5。
