High-affinity Cyclic Peptide Matriptase Inhibitors

Quimbar, Pedro; Malik, Uru; Sommerhoff, Christian P.; Kaas, Quentin; Chan, Lai Yue; Huang, Yen‐Hua; Grundhuber, Maresa; Dunse, Kerry; Craik, David J.; Anderson, Marilyn A.; Daly, Norelle L.

doi:10.1074/jbc.m113.460030

Cited by 130 publications

(139 citation statements)

References 46 publications

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“…In addition to wild-type KD1 (Ki = 310 pM ± 20 pM) (42,53), previous efforts have generated matriptase inhibitors based on constrained peptides (Ki = 830 ± 140 pM (40) and Ki = 290 ± 54 pM) (38) and antibodies (Ki = 720 pM) (39). Peptide and KD1-based inhibitors, as well as synthetic small molecule inhibitors (36,37), have short circulating half-lives and restricted molecular surface area for binding matriptase with high affinity.…”

Section: Discussionmentioning

confidence: 99%

“…Dose response plots were generated for each inhibitor ( Fig. S4A) and inhibition constant (Ki) values were determined using equation (1) and equation (2) as previously reported (36,38,40). Table 1 lists the resulting Ki value for each inhibitor construct and the number of functional Kunitz domains present.…”

Section: Kd1-kd2/1-fc Is a Potent And Selective Inhibitor Of Matriptasementioning

confidence: 99%

“…Alternative approaches to develop matriptase inhibitors include synthetic small molecules (36,37), peptides (38), monoclonal antibodies (39), and constrained peptide scaffolds (40). While each strategy generated molecules that bound to and inhibited matriptase activity, none address all of the reported therapeutic limitations.…”

Section: Introductionmentioning

confidence: 99%

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Engineering a potent inhibitor of matriptase from the natural hepatocyte growth factor activator inhibitor type-1 (HAI-1) protein

Mitchell

Kannan

Hunter

et al. 2018

Journal of Biological Chemistry

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Abstract:Dysregulated matriptase activity has been established as a key contributor to cancer progression through its activation of growth factors including the hepatocyte growth factor (HGF). Despite its critical role and prevalence in many human cancers, limitations to developing an effective matriptase inhibitor include weak binding affinity, poor selectivity, and short circulating half-life. We applied rational and combinatorial approaches to engineer a potent inhibitor based on the hepatocyte growth factor activator inhibitor type 1 (HAI-1), a natural matriptase inhibitor. The first Kunitz domain (KD1) of HAI-1 has been well established as a minimal matriptase binding and inhibition domain, while the second Kunitz domain (KD2) is inactive and involved in negative regulation. Here, we replaced the inactive KD2 domain of HAI-1 with an engineered chimeric variant of KD2/KD1 domains, and fused the resulting construct to an antibody Fc domain to increase valency and circulating serum half-life. The final protein variant contains 4 stoichiometric binding sites that we showed were needed to effectively inhibit matriptase with a Ki =70 ± 5 pM, an increase of 120-fold compared to the natural HAI-1 inhibitor, to our knowledge making it one of the most potent matriptase inhibitors identified to date. Furthermore, the engineered inhibitor demonstrates a protease selectivity profile similar to wild-type KD1 but distinct from HAI-1. It also inhibits activation of the natural pro-HGF substrate and matriptase expressed on cancer cells with at least an order of magnitude greater efficacy than KD1. ________________________________________ High expression and dysregulated activity of the type II, membrane-anchored serine protease matriptase in the local tumor environment has been shown to correlate with poor patient prognosis in many human cancers including breast, colorectal, pancreatic, cervical, and prostate cancers (1-10). This dysregulation is partly driven by the high proteolytic processing and turnover of pro-hepatocyte growth factor (Pro-HGF) (9, 11) to a form of HGF that activates its cognate receptor, c-Met (12) (Fig. 1A). Matriptase is also known to activate other proteases and growth factors including urokinase plasminogen activator (uPA) (11), pro-macrophage stimulating protein (Pro-MSP) (13), and platelet derived growth factor-D (PDGF-D) (14), all of which play key roles in cancer growth and metastasis. Furthermore, matriptase has been identified as a critical driver of other diseases, including iron overload disease (15) and osteoarthritis (16), and has been shown to activate the human airway influenza virus (H1N1) (17) and human immunodeficiency virus (HIV) (18 (19)(20)(21)(22)(23). The activity of matriptase is regulated in healthy tissue by the serine protease inhibitor hepatocyte growth factor activator inhibitor type-1 (HAI-1) (Fig. 1A). HAI-1 is primarily expressed on the surface of epithelial cells and naturally blocks the substrate-activating properties of matriptase (24)(25)(26), as well as other struc...

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Section: Discussionmentioning

confidence: 99%

Section: Kd1-kd2/1-fc Is a Potent And Selective Inhibitor Of Matriptasementioning

confidence: 99%

See 1 more Smart Citation

Engineering a potent inhibitor of matriptase from the natural hepatocyte growth factor activator inhibitor type-1 (HAI-1) protein

Mitchell

Kannan

Hunter

et al. 2018

Journal of Biological Chemistry

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“…Valinomycin, for example, is a cyclic dodecadepsipeptide produced by Streptomyces fulvissimus with potent antibacterial properties while also acting as a potassium-selective ionophore [19,20]. Enzyme inhibition is another pharmaceutical application that is associated with cyclic peptides, as shown by two examples of the Bowman-Birk class of protease inhibitors, Sunflower trypsin inhibitor-1 (SFTI-1) and Momordica cochinchinensis trypsin inhibitor-II (MCoTI-II) [21].…”

Section: Introductionmentioning

confidence: 99%

“…Stapled peptides suffer from a similar pitfall in the region of the peptide containing the stapled (cyclized) portion and also typically contain hydrocarbon (non-peptide-like) linkers that constitute the staple. Several mass spectrometric techniques, including collision induced dissociation (CID) [20][21][22][23][24][25], MS n methods [23,[25][26][27], electron capture dissociation (ECD) [28], and complexation strategies [29,30], have emerged as the most valuable tools to assist in the characterization of the sequences of non-linear peptides.…”

Section: Introductionmentioning

confidence: 99%

Exploitation of the Ornithine Effect Enhances Characterization of Stapled and Cyclic Peptides

Crittenden

Parker

Jenner

et al. 2016

J. Am. Soc. Mass Spectrom.

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Abstract. A method to facilitate the characterization of stapled or cyclic peptides is reported via an arginine-selective derivatization strategy coupled with MS/MS analysis. Arginine residues are converted to ornithine residues through a deguanidination reaction that installs a highly selectively cleavable site in peptides. Upon activation by CID or UVPD, the ornithine residue cyclizes to promote cleavage of the adjacent amide bond. This Arg-specific process offers a unique strategy for site-selective ring opening of stapled and cyclic peptides. Upon activation of each derivatized peptide, site-specific backbone cleavage at the ornithine residue results in two complementary products: the lactam ring-containing portion of the peptide and the aminecontaining portion. The deguanidination process not only provides a specific marker site that initiates fragmentation of the peptide but also offers a means to unlock the staple and differentiate isobaric stapled peptides.

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A Practical Guide to Structural Aspects of Macrocycles (NMR, X‐Ray, and Modeling)

Craik

Kaas

Wang

2017

Practical Medicinal Chemistry With Macrocycles

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High-affinity Cyclic Peptide Matriptase Inhibitors

Cited by 130 publications

References 46 publications

Engineering a potent inhibitor of matriptase from the natural hepatocyte growth factor activator inhibitor type-1 (HAI-1) protein

Engineering a potent inhibitor of matriptase from the natural hepatocyte growth factor activator inhibitor type-1 (HAI-1) protein

Exploitation of the Ornithine Effect Enhances Characterization of Stapled and Cyclic Peptides

A Practical Guide to Structural Aspects of Macrocycles (NMR, X‐Ray, and Modeling)

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