NMR-Based Structural Characterization of a Two-Disulfide-Bonded Analogue of the FXIIIa Inhibitor Tridegin: New Insights into Structure–Activity Relationships

Schmitz, Thomas; George, Ajay Abisheck Paul; Nubbemeyer, Britta; Bäuml, Charlotte A.; Steinmetzer, Torsten; Ohlenschläger, Oliver; Biswas, Arijit; Imhof, Diana

doi:10.3390/ijms22020880

Cited by 6 publications

(2 citation statements)

References 54 publications

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“…Overall, the sulfonated aromatic heparin mimetics platform, particularly inhibitor 16 , presents a unique alternative approach to modulate FXIIIa. − It appears to be very promising for providing molecules with potentially reduced bleeding risks. Furthermore, the use of specific molecules to perform molecular and cellular mechanistic studies may enhance our understanding of the role of FXIIIa in hemostasis.…”

Section: Discussionmentioning

confidence: 99%

Discovery of Heparin Mimetic, Potent, and Selective Inhibitors of Human Clotting Factor XIIIa

Vu,

Kar,

Goyal

et al. 2024

ACS Omega

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Factor XIIIa (FXIIIa) is a cysteine transglutaminase that catalyzes the last step in the coagulation process. An anionbinding site inhibition of FXIIIa is a paradigm-shifting strategy that may offer key advantages of controlled inhibition. Such an approach is likely to lead to novel FXIIIa inhibitors that do not carry bleeding risks. We previously reported a flavonoid trimer-based allosteric inhibitor of FXIIIa with moderate potency and selectivity. To further advance this approach, we evaluated a series of 27 variably sulfonated heparin mimetics against human FXIIIa. Only 13 molecules exhibited inhibitory activity at the highest concentration tested with IC 50 values of 2−286 μM. Specifically, inhibitor 16 demonstrated an IC 50 value of 2.4 ± 0.5 μM in a bisubstrate, fluorescence-based trans-glutamination assay. It also demonstrated a significant selectivity over other clotting factors including thrombin, factor Xa, and factor XIa as well as other cysteine enzymes including papain and tissue transglutaminase 2. Inhibitor 16 did not affect the viability of three human cell lines at a concentration that is 5-fold its FXIIIa-IC 50 . The molecule had a very weak effect on the activated partial thromboplastin time of human plasma at a concentration of >700 μM, further supporting its functional selectivity. Importantly, molecule 16 inhibited FXIIIa-mediated polymerization of fibrin(ogen) in a concentrationdependent manner as shown by the gel electrophoresis experiment. Michaelis−Menten kinetics revealed that the molecule competes with the Gln-donor protein substrate, i.e., dimethylcasein, but not with the Lys-donor small substrate, i.e., dansylcadaverine. Molecular modeling studies revealed that this type of molecule likely binds to an anion-binding site comprising the basic amino acids of Lys54, Lys61, Lys73, Lys156, and Arg244 among others. Overall, our work puts forward a new anion-binding site, selective, nontoxic, sulfonated heparin mimetic FXIIIa inhibitor 16 for further development as an effective and safer anticoagulant.

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Section: Discussionmentioning

confidence: 99%

Discovery of Heparin Mimetic, Potent, and Selective Inhibitors of Human Clotting Factor XIIIa

Vu,

Kar,

Goyal

et al. 2024

ACS Omega

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“…Tridegin, a small peptide inhibitor purified from the salivary gland extract of the giant Amazon leech Haementeria ghilianii, inhibits both plasma and platelet FXIIIa without interfering with the enzymatic activity of thrombin or Factor Xa [214]. Analogues of this peptidic inhibitor offer insight into the mechanism of action and have potential as lead structures for development [215]. The novel inhibitors with a cis-bisamido epoxides pharmacore were shown to have an improved potency compared to a natural product inhibitor, cerulenin, although still lacked selectivity for FXIII over transglutaminase 2 [216].…”

Section: Fxiii-a Replacement Therapy and Utility As A Drug Targetmentioning

confidence: 99%

Factor XIII-A: An Indispensable “Factor” in Haemostasis and Wound Healing

Alshehri

Whyte

Mutch

2021

IJMS

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Factor XIII (FXIII) is a transglutaminase enzyme that catalyses the formation of ε-(γ-glutamyl)lysyl isopeptide bonds into protein substrates. The plasma form, FXIIIA2B2, has an established function in haemostasis, with fibrin being its principal substrate. A deficiency in FXIII manifests as a severe bleeding diathesis emphasising its crucial role in this pathway. The FXIII-A gene (F13A1) is expressed in cells of bone marrow and mesenchymal lineage. The cellular form, a homodimer of the A subunits denoted FXIII-A, was perceived to remain intracellular, due to the lack of a classical signal peptide for its release. It is now apparent that FXIII-A can be externalised from cells, by an as yet unknown mechanism. Thus, three pools of FXIII-A exist within the circulation: plasma where it circulates in complex with the inhibitory FXIII-B subunits, and the cellular form encased within platelets and monocytes/macrophages. The abundance of this transglutaminase in different forms and locations in the vasculature reflect the complex and crucial roles of this enzyme in physiological processes. Herein, we examine the significance of these pools of FXIII-A in different settings and the evidence to date to support their function in haemostasis and wound healing.

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Innovative Pharmaceutical Stabilization and Formulation Processes for Protein Drugs and Their Impact on Sequence and Structure. Part: Analytics

Elsayed,

Kühl,

Imhof

2024

Dispersity, Structure and Phase Changes of Proteins and Bio Agglomerates in Biotechnological Processes

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NMR-Based Structural Characterization of a Two-Disulfide-Bonded Analogue of the FXIIIa Inhibitor Tridegin: New Insights into Structure–Activity Relationships

Cited by 6 publications

References 54 publications

Discovery of Heparin Mimetic, Potent, and Selective Inhibitors of Human Clotting Factor XIIIa

Discovery of Heparin Mimetic, Potent, and Selective Inhibitors of Human Clotting Factor XIIIa

Factor XIII-A: An Indispensable “Factor” in Haemostasis and Wound Healing

Innovative Pharmaceutical Stabilization and Formulation Processes for Protein Drugs and Their Impact on Sequence and Structure. Part: Analytics

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