Visn. Nac. Akad. Nauk Ukr. 2019. (4):50-56
https://doi.org/10.15407/visn2019.04.050
V.O. Chernyshenko
Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv
MECHANISMS OF INTRAVASCULAR AND EXTRAVASCULAR BLOOD CLOTTING: FUNDAMENTAL STUDIES FOR CLINICAL PRACTICE
According to the materials of scientific report at the meeting of the Presidium of NAS of Ukraine, February 27, 2019
Haemostasis is the unique system of human organism that support blood flow in vessels and the termination of blood loss after injury. The dynamic balance of haemostasis is provided by the interplay of several enzymatic systems that all are targeted directly or indirectly on fibrinogen. Fibrinogen is soluble protein of blood that when activated by thrombin converts to fibrin that is able to form polymeric fibrin which is the core of thrombus. To study the structure and functions of fibrin several molecular instruments are used, among them are monoclonal antibodies, synthetic peptides, low-molecular weight compounds and specific proteases.
Monoclonal antibody I-5А specific to fibrinogen fragment Аα509-610 is shown to be an effective inhibitor of lateral association of protofibrils during fibrin polymerization. Thus we assume the presence of some previously unknown site of lateral association of protofibrils within the αC-region of molecule. Also inhibitory antibody I-5А or its recombinant single chain analogue (scFv) can be assumed as the prospective agent for the inhibition of thrombus formation in vivo.
We also found the residues of fibrinogen molecule that participate in both stages of fibrin polymerization process: Aα-91-103, Bβ-125-135 and γ-69-77. The synthetic peptides that mimic these residues together or separately can be used for prevention of thrombus formation.
Finally, unique calix[4]arene C-145 with strong antithrombotic activity is found. We demonstrate that C-145 directly blocked A:a knob-hole interactions during fibrin polymerization thus preventing the formation of three-dimensional fibrin network that is the core of thrombus. Such unique properties allow assuming the possible use of C-145 in antithrombotic therapy. In vivo studies demonstrated that C-145 being introduced to laboratory mice per orally acts as effective anticoagulant agent.
Apart of the prevention of intravascular coagulation we also focus on the initiation of extravascular clotting that is one of the crucial points for the prevention of blood loss after traumas or in surgery. Direct comparison in the model of massive blood loss demonstrated that developed haemostatic agent was more effective for blood loss prevention then commercially available Celox (UK).
Keywords: blood circulation, thrombosis, bleeding, antibodies, peptides, calix[4]arenes, haemostatics.
Language of article: ukrainian
REFERENCES
1. Belitser V.A., Varetska T.V., Manjakov V.Ph. On the Model of the Fibrinogen Molecule. Conclusive Stages of Fibrin Polymerization. Thromb. Research. 1973. 2: 567. https://doi.org/10.1016/0049-3848(73)90008-X
2. Doolittle R.F. Structural basis of the fibrinogen-fibrin transformation: contributions from X-ray crystallography. Blood Rev. 2003. 1: 33. https://doi.org/10.1016/S0268-960X(02)00060-7
3. Medved L., Weisel J.W. Recommendations for nomenclature on fibrinogen and fibrin. J. Thromb. Haemost. 2009. 7(2): 355. https://doi.org/10.1111/j.1538-7836.2008.03242.x
4. Lugovskoi E.V., Makogonenko E.M., Komisarenko S.V. Molecular mechanisms of formation and degradation of fibrin. (Kyiv: Naukova Dumka, 2013).
5. Lugovskoy E.V., Gritsenko P.G., Kapustianenko L.G. et al. Functional role of Bβ-chain N-terminal fragment in the fibrin polymerization process. FEBS J. 2007. 274: 4540. https://doi.org/10.1111/j.1742-4658.2007.05983.x
6. Springer T.A., Zhu J., Xiao T. Structural basis for distinctive recognition of fibrinogen gammaC peptide by the platelet integrin alphaIIbbeta3. J. Cell Biol. 2008. 182(4): 791. https://doi.org/10.1083/jcb.200801146
7. Lugovskoi E.V., Makogonenko E.M., Chudnovets S.G. et al. The study of fibrin polymerization with monoclonal antibodies. Biomed. Sci. 1991. 2(3): 249.
8. Lugovskоi E.V., Kolesnikova I.N., Platonova T.N. et al. Simultaneous quantification of soluble fibrin and D-dimer in blood plasma for the assessment of the threat of thrombosis. Klin. Med. (Mosk.). 2013. 91(11): 38.
9. Urvant L., Makogonenko Y., Pozniak T. et al. On the role of αc-regions of fibrin in the self-assembly and lateral association of protofibrils. FEBS J. 2013. 280(s1): 489. https://doi.org/10.1111/febs.12340
10. Pozniak T.A., Urvant L.P., Gritsenko P.G. et al. Inhibition of fibrin polymerization by synthetic peptides corresponding to Aα195-205 and γ69-77 sites of fibrin molecule. Ukr. Biochem. J. 2014. 86(4): 119. http://doi.org/10.15407/ubj86.04.119
11. Lugovskoy E.V., Gritsenko P.G., Koshel T.A. et al. Calix[4]arene methylenebisphosphonic acids as inhibitors of fibrin polymerization. FEBS J. 2011. 278(8): 1244. https://doi.org/10.1111/j.1742-4658.2011.08045.x
12. Komisarenko S.V., Lugovskoi E.V., Nikolaev V.G. et al. Combined haemostatic agent for the prevention of blood loss in particular at hemophilia. Inventory patent. 2017. 19. 114356.