In Vivo Imaging of MMP‐13 Activity Using a Specific Polymer‐FRET Peptide Conjugate Detects Early Osteoarthritis and Inhibitor Efficacy

Abstract

Imaging early molecular changes in osteoarthritic (OA) joints is instrumental for the development of disease-modifying drugs. To this end, a fluorescent resonance energy transfer-based peptide probe that is cleavable by matrix metalloproteinase 13 (MMP-13) has been developed. This protease degrades type II collagen, a major matrix component of cartilage. The probe exhibits high catalytic efficiency (k cat /K M = 6.5 × 10 5 m −1 s −1 ) and high selectivity for MMP-13 over a set of nine MMPs. To achieve optimal in vivo pharmacokinetics and tissue penetration, the probe has been further conjugated to a linear l-polyglutamate chain of 30 kDa. The conjugate detects early biochemical events that occur in a surgically induced murine model of OA before major histological changes. The nanometric probe is suitable for the monitoring of in vivo efficacy of an orally bioavailable MMP-13 inhibitor, which effectively blocks cartilage degradation during the development of OA. This new polymer-probe can therefore be a useful tool in detecting early OA, disease progression, and in developing MMP-13-based disease-modifying drugs for OA.

MMP-13 Enzyme and pH Responsive Theranostic Nanoplatform for Osteoarthritis

“…Here, a specific collagen type II targeting peptide (Coll-II α1 chain-binding peptide -CollB) with the sequence of WRYGRL [16][17][18] was grafted onto PPL to form cartilage targeting PPL (C-PPL). In parallel, PPL was conjugated with a specific peptide substrate of MMP-13 enzyme (H 2 N-GPLGVRGC-SH) [19,20] labeled with a fluorescence dye (Cy5.5). Further, black hole quencher-3 (BHQ-3) that can quench Cy5.5 fluorescence was coupled via amide reaction to obtain MMP-13 responsive and pH sensitive polymer MR-Cy5.5-BHQ-3-PPL (MR-PPL).…”

Application of FRET Biosensors in Mechanobiology and Mechanopharmacological Screening

“…Similar cleavage-based FRET reporters have been constructed for MMP-2 (Yang et al, 2007;Lee et al, 2020), MMP-9 (Stawarski et al, 2014), MMP-11 (Meyer and Rademann, 2012), MMP-12 (Lim et al, 2014), and MMP-13 (Lim et al, 2014;Duro-Castano et al, 2018). Interestingly, a dual-FRET biosensor was developed for simultaneous imaging of MMP-2 and caspase-3 activities, in which the sequential cleavage by MMP-2 and caspase-3 increased the donor emission intensity in a stepwise fashion (Li et al, 2015).…”

The Impact of Chemical Biology on Drug Discovery

“…As one approach to address this issue, we developed a concept for signal localization relying on lipidation that allowed specific detection of tumor tissue . We were also able to apply the concept of activity‐based drugs to examine emerging treatment concepts of osteoarthritis in in vivo models, namely by detecting the effects of a selective, orally bioavailable MMP‐13 inhibitor on cartilage degradation with a novel FRET based probe . Fluorescence, despite its limited spatial resolution, can prove to be a valuable tool for evaluation of compound effects not only in cells but also in small rodents.…”