Design and chemical syntheses of potent matriptase‐2 inhibitors based on trypsin inhibitor SFTI‐1 isolated from sunflower seeds

Gitlin-Domagalska, Agata; Dębowski, Dawid; Łęgowska, Anna; Stirnberg, Marit; Okońska, Joanna; Gütschow, Michael; Rolka, Krzysztof

doi:10.1002/bip.23031

Cited by 14 publications

(11 citation statements)

References 64 publications

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“…The proteolytic activity of MT2 has been identified as a target for the treatment of iron overload disorders by application of specific inhibitors to block the function of MT2 that is predicted to elevate hepcidin expression. 53,58,59 Our data suggest that such an approach might not be able to achieve this goal. On the basis of the findings in this and previous studies, we propose a model in which Mt2 binds to substrates on the plasma membrane through the CUB and catalytic domains, followed by rapid removal of substrates from the cells via its protease activity.…”

Section: Discussionmentioning

confidence: 78%

The ectodomain of matriptase-2 plays an important nonproteolytic role in suppressing hepcidin expression in mice

Enns

Jue

Zhang

2020

Blood

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Matriptase-2 (MT2), encoded by TMPRSS6, is a membrane-anchored serine protease, which plays a key role in suppressing hepatic hepcidin expression. MT2 is synthesized as a zymogen and undergoes autocleavage for activation. Previous studies suggest that MT2 suppresses hepcidin by cleaving hemojuvelin (HJV) and other components of the BMP-signaling pathway. However, the underlying mechanism is still debatable. Here we dissected the contributions of the non-proteolytic and proteolytic activities of Mt2 by taking advantage of Mt2 mutants and Tmprss6-/- mice. Studies of the protease-dead full-length Mt2 (Mt2S762A) and the truncated Mt2 that lacks the catalytic domain (Mt2mask) indicate that the catalytic domain, but not its proteolytic activity, was required for Mt2 to suppress hepcidin expression. This process was likely accomplished by the binding of Mt2 ectodomain to Hjv and Hfe. We found that Mt2 specifically cleaved the key components of the hepcidin induction pathway including Hjv, Alk3, ActRIIA, and Hfe when overexpressed in hepatoma cells. Nevertheless, studies of a murine IRIDA-causing mutant (Mt2I286F) in the CUB domain indicate that Mt2I286F can be activated but it displayed a largely compromised ability to suppress hepcidin expression. Co-immunoprecipitation analysis revealed that Mt2I286F, but not Mt2S762A, had reduced interactions with Hjv, ActRIIA, and Hfe. Additionally, increased expression of the serine protease inhibitor Hai-2 in the liver failed to alter hepcidin. Together these observations support the idea that the substrate interaction with Mt2 plays a determinant role, and suggest that the proteolytic activity is not an appropriate target to modulate the function of MT2 for clinical applications.

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Section: Discussionmentioning

confidence: 78%

The ectodomain of matriptase-2 plays an important nonproteolytic role in suppressing hepcidin expression in mice

Enns

Jue

Zhang

2020

Blood

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“…The 14-mer cyclic sunflower trypsin inhibitor-1 (SFTI-1 ( 72 ), trypsin K i = 17 pM) is the smallest member of the Bowman–Birk family of serine protease inhibitors (Table ). − Its head to tail ring structure is stabilized by an intramolecular disulfide bridge and hydrogen bond network. , The SFTI-1-binding traps the target protease in a futile cycle of cleavage and religation of the scissile P1–P1′ peptide bond between Lys5-Ser6 according to a Laskowski mechanism. , Based on the SFTI-1 scaffold, several highly potent and selective inhibitors of numerous serine proteases such as thrombin, chymotrypsin, matriptase-1/-2, , cathepsin G, various kallikrein-related peptidases, ,, and furin have been developed.…”

Section: Synthetic Active Site-directed Inhibitorsmentioning

confidence: 99%

“…191,192 The SFTI-1-binding traps the target protease in a futile cycle of cleavage and religation of the scissile P1−P1′ peptide bond between Lys5-Ser6 according to a Laskowski mechanism. 193,194 Based on the SFTI-1 scaffold, several highly potent and selective inhibitors of numerous serine proteases such as thrombin, 190 chymotrypsin, 195 matriptase-1/-2, 196,197 cathepsin G, 198 various kallikrein-related peptidases, 190,199,200 and furin 201 have been developed.…”

Section: Synthetic Active Site-directed Inhibitorsmentioning

confidence: 99%

Fibrinolysis Inhibitors: Potential Drugs for the Treatment and Prevention of Bleeding

et al. 2019

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Hyperfibrinolytic situations can lead to lifethreatening bleeding, especially during cardiac surgery. The approved antifibrinolytic agents such as tranexamic acid, εaminocaproic acid, 4-aminomethylbenzoic acid, and aprotinin were developed in the 1960s without the structural insight of their respective targets. Crystal structures of the main antifibrinolytic targets, the lysine binding sites on plasminogen's kringle domains, and plasmin's serine protease domain greatly contributed to the structure-based drug design of novel inhibitor classes. Two series of ligands targeting the lysine binding sites have been recently described, which are more potent than the most-widely used antifibrinolytic agent, tranexamic acid. Furthermore, four types of promising active site inhibitors of plasmin have been developed: tranexamic acid conjugates targeting the S1 pocket and primed sites, substrate-analogue linear homopiperidylalanine-containing 4amidinobenzylamide derivatives, macrocyclic inhibitors addressing nonprimed binding regions, and bicyclic 14-mer SFTI-1 analogues blocking both, primed and nonprimed binding sites of plasmin. Furthermore, several allosteric plasmin inhibitors based on heparin mimetics have been developed.

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“…Its cyclic peptide backbone and bisecting disulfide bond make SFTI-1 highly stable and thus an attractive scaffold for design of therapeutic compounds targeting serine proteases, as well as cell surface receptors and protein–protein interactions . SFTI-1 has been used to engineer potent and/or selective inhibitors of a number of serine proteases, including thrombin, chymotrypsin, matriptase-1/-2, , cathepsin G, and several kallikrein-related peptidases, ,, highlighting the versatility of the scaffold.…”

Section: Introductionmentioning

confidence: 99%

Highly Potent and Selective Plasmin Inhibitors Based on the Sunflower Trypsin Inhibitor-1 Scaffold Attenuate Fibrinolysis in Plasma

Swedberg

Mahatmanto

et al. 2018

J. Med. Chem.

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Antifibrinolytic drugs provide important pharmacological interventions to reduce morbidity and mortality from excessive bleeding during surgery and after trauma. Current drugs used for inhibiting the dissolution of fibrin, the main structural component of blood clots, are associated with adverse events due to lack of potency, high doses, and nonselective inhibition mechanisms. These drawbacks warrant the development of a new generation of highly potent and selective fibrinolysis inhibitors. Here, we use the 14-amino acid backbone-cyclic sunflower trypsin inhibitor-1 scaffold to design a highly potent (K i = 0.05 nM) inhibitor of the primary serine protease in fibrinolysis, plasmin. This compound displays a million-fold selectivity over other serine proteases in blood, inhibits fibrinolysis in plasma more effectively than the gold-standard therapeutic inhibitor aprotinin, and is a promising candidate for development of highly specific fibrinolysis inhibitors with reduced side effects.

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Design and chemical syntheses of potent matriptase‐2 inhibitors based on trypsin inhibitor SFTI‐1 isolated from sunflower seeds

Cited by 14 publications

References 64 publications

The ectodomain of matriptase-2 plays an important nonproteolytic role in suppressing hepcidin expression in mice

The ectodomain of matriptase-2 plays an important nonproteolytic role in suppressing hepcidin expression in mice

Fibrinolysis Inhibitors: Potential Drugs for the Treatment and Prevention of Bleeding

Highly Potent and Selective Plasmin Inhibitors Based on the Sunflower Trypsin Inhibitor-1 Scaffold Attenuate Fibrinolysis in Plasma

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