Generation of Highly Selective MMP Antibody Inhibitors

Nam, Dong Hyun; Ge, Xin

doi:10.1007/978-1-4939-7595-2_26

Cited by 8 publications

(8 citation statements)

References 35 publications

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“…As previously mentioned, monoclonal antibodies [120] are also among several additional experimental MMP/ADAMs inhibitors undergoing pre-clinical testing in various inflammatory disorders and cancer. These include, MMP-14 [121], MMP-9 [122], and, ADAM17 [123].…”

Section: In Vitro Cell Cultures Appear To Be Integral For Assessing the Action Of Mmp Inhibitorsmentioning

confidence: 99%

Inhibition of MMPs and ADAM/ADAMTS

Malemud

2019

Biochemical Pharmacology

141

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Matrix metalloproteinases (MMPs), A Disintegrin and Metalloproteinase (ADAM) and A Disintegrin and Metalloproteinase with Thrombospondin Motif (ADAMTS) are zinc-dependent endopeptidases that play a critical role in the destruction of extracellular matrix proteins and, the shedding of membrane-bound receptor molecules in various forms of arthritis and other diseases. Under normal conditions, MMP, ADAM and ADAMTS gene expression aids in the maintenance of homeostasis. However, in inflamed synovial joints characteristic of rheumatoid arthritis and osteoarthritis. MMP, ADAM and ADAMTS production is greatly increased under the influence of pro-inflammatory cytokines. Analyses based on medicinal chemistry strategies designed to directly inhibit the activity of MMPs have been largely unsuccessful when these MMP inhibitors were employed in animal models of rheumatoid arthritis and osteoarthritis. This is despite the fact that these MMP inhibitors were largely able to suppress pro-inflammatory cytokine-induced MMP production in vitro. A focus on ADAM and ADAMTS inhibitors has also been pursued. Thus, recent progress has identified the "sheddase" activity of ADAMs as a viable target and the development of GW280264X is an experimental ADAM17 inhibitor. Of note, a monoclonal antibody, GLPG1972, developed as an ADAMTS-5 inhibitor, entered a Phase I OA clinical trial. However, the failure of many of these previously developed inhibitors to move beyond the preclinical testing phase has required that novel strategies be developed that are designed to suppress both MMP, ADAM and ADAMTS production and activity.

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Section: In Vitro Cell Cultures Appear To Be Integral For Assessing the Action Of Mmp Inhibitorsmentioning

confidence: 99%

Inhibition of MMPs and ADAM/ADAMTS

Malemud

2019

Biochemical Pharmacology

141

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“…We have used several strategies to identify specific inhibitory antibodies of uPA. Based on previously identified potent antibody inhibitors against serine proteases [26,27], and matrix metallopeptidases (MMPs) [12], where the main driver of inhibition is the long CDRH3 loop inserted into the protease active site [27,28], we designed a set of synthetic antibody libraries based on the long CDRH3 loops of two potent MTSP-1 inhibitory Fabs (Unpublished results). Several uPA antibodies were identified using these biased antibody libraries but none showed an inhibitory effect, and some behaved as substrates of urokinase ( Figure Supplementary Figure 3).…”

Section: Discussionmentioning

confidence: 99%

“…Our results for protease production and identification of rAbs to MT-SP1 (aka, matriptase), and urokinase plasminogen activator (uPA) showed that conformationally selective inhibitory antibodies can be identified from a naïve human Fab library displayed on bacteriophage. Several inhibitory antibodies have been developed to other proteases including HGFA [10], Factor XIa [11], MMP9 [12] and β-tryptase [13]. Another advantage of rAbs is that the antibody sequence of the binder allows for in vitro affinity maturation, mimicking an immune response, to select Abs with higher affinity and lower off-rate [14].…”

Section: Introductionmentioning

confidence: 99%

Structure of an affinity-matured inhibitory recombinant fab against urokinase plasminogen activator reveals basis of potency and specificity

Sevillano

Bohn

Zimanyi

et al. 2021

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

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Affinity maturation of U33, a recombinant Fab inhibitor of uPA, was used to improve the affinity and the inhibitory effect compared to the parental Fab. Arginine scanning of the six CDR loops of U33 was done to identify initial binding determinants since uPA prefers arginine in its primary substrate binding pocket. Two CDR loops were selected to create an engineered affinity maturation library of U33 that was diversified around ArgL91 (CDR L3) and ArgH52 (CDR H2). Biopanning of the randomized U33 library under stringent conditions resulted in eight Fabs with improved binding properties. One of the most potent inhibitors, AB2, exhibited a 13fold decrease in IC50 when compared to U33 largely due to a decrease in its off rate. To identify contributions of interfacial residues that might undergo structural rearrangement upon interface formation we used X-ray footprinting and mass spectrometry (XFMS). Four residues showed a pronounced decrease in solvent accessibility, and their clustering suggests that AB2 targets the active site and also engages residues in an adjacent pocket unique to human uPA. The 2.9 Å resolution crystal structure of AB2-bound to uPA shows a binding mode in which the CDR L1 loop inserts into the active site cleft and acts as a determinant of inhibition. The selectivity determinant of this binding mode is unlike previously identified inhibitory Fabs against uPA related serine proteases, MTSP-1, HGFA and FXIa. CDRs H2 and L3 loops aid in interface formation and provide critical saltbridges to remodel loops surrounding the active site of uPA providing specificity and further evidence that antibodies can be potent and selective inhibitors of proteolytic enzymes.

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“…A monoclonal antibody against MMP-14 selectively blocks proMMP-2 activation which is required for lymphangiogenesis in vitro and ex vivo [ 443 ]. In preclinical models, several monoclonal antibodies against MMP-9 [ 444 , 445 ] and MMP-14 [ 446 ] initially appeared promising, but the generation of highly selective MMP antibody inhibitors remains a desideratum [ 447 ]. Phage display library-derived peptides against the MT loop of MMP-14, such as HS7, which may be useful in the early diagnosis of tumors and for the development of peptide-mediated drugs [ 448 ].…”

Section: Translational Assessment and Future Prospectmentioning

confidence: 99%

Hold on or Cut? Integrin- and MMP-Mediated Cell–Matrix Interactions in the Tumor Microenvironment

Niland

Eble

2020

IJMS

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The tumor microenvironment (TME) has become the focus of interest in cancer research and treatment. It includes the extracellular matrix (ECM) and ECM-modifying enzymes that are secreted by cancer and neighboring cells. The ECM serves both to anchor the tumor cells embedded in it and as a means of communication between the various cellular and non-cellular components of the TME. The cells of the TME modify their surrounding cancer-characteristic ECM. This in turn provides feedback to them via cellular receptors, thereby regulating, together with cytokines and exosomes, differentiation processes as well as tumor progression and spread. Matrix remodeling is accomplished by altering the repertoire of ECM components and by biophysical changes in stiffness and tension caused by ECM-crosslinking and ECM-degrading enzymes, in particular matrix metalloproteinases (MMPs). These can degrade ECM barriers or, by partial proteolysis, release soluble ECM fragments called matrikines, which influence cells inside and outside the TME. This review examines the changes in the ECM of the TME and the interaction between cells and the ECM, with a particular focus on MMPs.

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Generation of Highly Selective MMP Antibody Inhibitors

Cited by 8 publications

References 35 publications

Inhibition of MMPs and ADAM/ADAMTS

Inhibition of MMPs and ADAM/ADAMTS

Structure of an affinity-matured inhibitory recombinant fab against urokinase plasminogen activator reveals basis of potency and specificity

Hold on or Cut? Integrin- and MMP-Mediated Cell–Matrix Interactions in the Tumor Microenvironment

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