VITPOR AI, A Coagulation Factor XIIa Inhibitor from Porphyra yezoensis: In Vivo Mode of Action and Assessment of Platelet Function Analysis

Author(s): Kalkooru L. Venkatraman, Azeemullah A. Syed, Parimelazhagan Indumathi, Alka Mehta*

Journal Name: Protein & Peptide Letters

Volume 27 , Issue 3 , 2020

Become EABM
Become Reviewer
Call for Editor

Graphical Abstract:


Background: Thrombosis represents as the prime contributor to the burden of diseases, worldwide. Conventional anticoagulants for thrombosis therapy have a common bleeding side effect. Bioactive peptides are studied to be an effective alternative for currently available therapeutic drugs.

Objective: In this study, VITPOR AI peptide, a previously reported coagulation FXIIa inhibitor from Nori (Porphyra yezoensis), was assessed for its inhibitory activity against FXIIa and its in vivo mode of action.

Methods: In vivo efficacy as well as the antithrombotic property of the peptide was evaluated in mice model by ex vivo activated Partial Thromboplastin Time assay, tail transection model and whole blood clotting time. The enzyme kinetics was studied using chromogenic substrate assay.

Results: The kinetic behaviour of VITPOR AI showed that the peptide is a competitive inhibitor of FXIIa. Peptide showed significant inhibition of platelet adhesion and aggregation. VITPOR AI exhibited significant antithrombotic activity. Furthermore, ex vivo activated Partial Thromboplastin Time assay revealed that VITPOR AI exhibited potent anticoagulant activity in vivo. Tail bleeding assay revealed that the peptide did not prolong bleeding time in mice even at a higher dose of 5 mg/kg. Cytotoxicity studies of the peptide against human blood leukocytes indicated the safety of the peptide.

Conclusion: VITPOR AI could be prospected as a potent anticoagulant with Factor XIIa inhibition, antiplatelet aggregation and antithrombotic activity. It was also studied to have no bleeding side effect.

Keywords: Anticoagulant peptide, factor XIIa inhibitor, Porphyra yezoensis hydrolysate, platelet aggregation and adhesion, leukocytes.

Davie, E.W.; Ratnoff, O.D. Waterfall sequence for intrinsic blood clotting. Science, 1964, 145(3638), 1310-1312.
[] [PMID: 14173416]
Yarovaya, G.A.; Blokhina, T.B.; Neshkova, E.A. Contact system. New concepts on activation mechanisms and bioregulatory functions. Biochemistry (Mosc.), 2002, 67(1), 13-24.
[] [PMID: 11841336]
Mackman, N. Triggers, targets and treatments for thrombosis. Nature, 2008, 451(7181), 914-918.
[] [PMID: 18288180]
Kenne, E.; Nickel, K.F.; Long, A.T.; Fuchs, T.A.; Stavrou, E.X.; Stahl, F.R.; Renné, T. Factor XII: A novel target for safe prevention of thrombosis and inflammation. J. Intern. Med., 2015, 278(6), 571-585.
[] [PMID: 26373901]
Decrem, Y.; Rath, G.; Blasioli, V.; Cauchie, P.; Robert, S.; Beaufays, J.; Frère, J.M.; Feron, O.; Dogné, J.M.; Dessy, C.; Vanhamme, L.; Godfroid, E. Ir-CPI, a coagulation contact phase inhibitor from the tick Ixodes ricinus, inhibits thrombus formation without impairing hemostasis. J. Exp. Med., 2009, 206(11), 2381-2395.
[] [PMID: 19808248]
Pauer, H.U.; Renné, T.; Hemmerlein, B.; Legler, T.; Fritzlar, S.; Adham, I.; Müller-Esterl, W.; Emons, G.; Sancken, U.; Engel, W.; Burfeind, P. Targeted deletion of murine coagulation factor XII gene-a model for contact phase activation in vivo. Thromb. Haemost., 2004, 92(3), 503-508.
[] [PMID: 15351846]
Iwaki, T.; Castellino, F.J. Plasma levels of bradykinin are suppressed in factor XII-deficient mice. Thromb. Haemost., 2006, 95(6), 1003-1010.
[] [PMID: 16732380]
Renné, T.; Schmaier, A.H.; Nickel, K.F.; Blombäck, M.; Maas, C. In vivo roles of factor XII. Blood, 2012, 120(22), 4296-4303.
[] [PMID: 22993391]
Rajapakse, N.; Jung, W.K.; Mendis, E.; Moon, S.H.; Kim, S.K. A novel anticoagulant purified from fish protein hydrolysate inhibits factor XIIa and platelet aggregation. Life Sci., 2005, 76(22), 2607-2619.
[] [PMID: 15769484]
Jo, H.Y.; Jung, W.K.; Kim, S.K. Purification and characterization of a novel anticoagulant peptide from marine echiuroid worm, Urechis unicinctus. Process Biochem., 2008, 43(2), 179-184.
Koyama, T.; Noguchi, K.; Aniya, Y.; Sakanashi, M. Analysis for sites of anticoagulant action of plancinin, a new anticoagulant peptide isolated from the starfish Acanthaster planci. Gen. Pharmacol., 1998, 31(2), 277-282.
Karasudani, I.; Koyama, T.; Nakandakari, S.; Aniya, Y. Purification of anticoagulant factor from the spine venom of the crown-of thorns starfish Acanthaster Planci. Toxicon, 1996, 34(8), 871-879.
Jung, W.; Kim, S. Isolation and characterisation of an anticoagulant oligopeptide from blue mussel Mytilus edulis. Food Chem., 2009, 117, 687-692.
[ hem.2009.04.077]
Waxman, L.; Smith, D.E.; Arcuri, K.E.; Vlasuk, G.P. Tick Anticoagulant Peptide (TAP) is a novel inhibitor of blood coagulation factor Xa. Science, 1990, 248(4955), 593-596.
[] [PMID: 2333510]
Feng, L.; Maolin, T.; Meiling, Q.; Fengjiao, F.; Hui, C.; Wei, S.; Ming, D. Thrombin inhibitory peptides derived from Mytilus edulis proteins: Identification, molecular docking and in silico prediction of toxicity. Eur. Food Res. Technol., 2018, 244(2), 207-217.
Syed, A.A.; Mehta, A. Target specific anticoagulant peptides: A Review. Int. J. Pept. Res. Ther., 2018, 24(1), 1-12.
Kong, Y.; Shao, Y.; Chen, H.; Ming, X.; Wang, J.B.; Li, Z.Y.; Wei, J.F. A novel factor Xa-inhibiting peptide from centipede’s venom. Int. J. Pept. Res. Ther., 2013, 19(4), 303-311.
[] [PMID: 24273471]
Thakur, R.; Kumar, A.; Bose, B.; Panda, D.; Saikia, D.; Chattopadhyay, P.; Mukherjee, A.K. A new peptide (Ruviprase) purified from the venom of Daboia russelii russelii shows potent anticoagulant activity via non-enzymatic inhibition of thrombin and factor Xa. Biochimie, 2014, 105, 149-158.
[] [PMID: 25038567]
Chen, M.; Ye, X.; Ming, X.; Chen, Y.; Wang, Y.; Su, X.; Su, W.; Kong, Y. A novel direct factor Xa inhibitory peptide with anti-platelet aggregation activity from Agkistrodon acutus venom hydrolysates. Sci. Rep., 2015, 5, 10846.
[] [PMID: 26035670]
Indumathi, P.; Mehta, A. A novel anticoagulant peptide from the Nori hydrolysate. J. Funct. Foods, 2016, 20, 606-617.
Syed, A.A.; Venkatraman, K.L.; Mehta, A. An anticoagulant peptide from Porphyra yezoensis inhibits the activity of factor XIIa: In vitro and in silico analysis. J. Mol. Graph. Model., 2019, 89, 225-233.
[] [PMID: 30921556]
Ishimaru, Y.; Gomez, E.A.; Zhang, F.; Martini-Robles, L.; Iwata, H.; Sakurai, T.; Katakura, K.; Hashiguchi, Y.; Kato, H. Dimiconin, a novel coagulation inhibitor from the kissing bug, Triatoma dimidiata, a vector of Chagas disease. J. Exp. Biol., 2012, 215(Pt 20), 3597-3602.
[] [PMID: 22771751]
Bendar, B.C.; Condra, R.J. Platelet aggregation monitored in a 96-well microplate reader is useful for evaluation of platelet agonists and antagonists. Thromb. Res., 1995, 77, 453-463.
Bellavite, P.G.; Andrioli, P.; Guzzo, P.; Arigliano, S.; Chirumbolo, F.; Manzato, C. Santonastaso A colorimetric method for the measurement of platelet adhesion in microtiter plates. Anal. Biochem, 1994, 216, 444-450.15.
Yang, W.G.; Wang, Z.; Xu, S.Y. A new method for determination of antithrombotic activity of egg white protein hydrolysate by microplate reader. Chin. Chem. Lett., 2007, 18(4), 449-451.
Zhang, S.B.; Wang, Z.; Xu, S.Y. Antioxidant and antithrombotic activities of rapeseed peptides. J. Am. Oil Chem. Soc., 2008, 85(6), 521-527.
Kruisbeek, A.M.E. Proliferative assays for T cells, In: Protocols in Immunology, Coligan, E.; Krusibeek, A.M.; Margnlies, D.H.; Shevach, E.M.; Strober, W.; Eds.; Current, John Wiley & Sons: New York. 2004, 60, p. (10)3.12.1-3.12.7.
Green, L.M.; Reade, J.L.; Wade, C.F. Rapid colorimetric assay for cell viability: application to the quantification of cytotoxic and growth inhibitory lymphokines. J. Immunol. Methods, 1984, 70, 257-268.
[ (84)90190-X]
Huang, J.; Wang, S.; Luo, X.; Xie, Y.; Shi, X. Cinnamaldehyde reduction of platelet aggregation and thrombosis in rodents. Thromb. Res., 2007, 119(3), 337-342.
[] [PMID: 16626787]
2Bou-Gharios, G.; Ponticos, M.; Rajkumar, V.; Abraham, D. Extracellular matrix in vascular networks. Cell Prolif., 2004, 37, 207-220.
Cohen, M. Platelet glycoprotein IIb/IIIa receptor inhibitors in coronary artery disease. Ann. Intern. Med., 1996, 124(9), 843-844.
[] [PMID: 8610955]
Pedicord, D.L.; Seiffert, D.; Blat, Y. Feedback activation of factor XI by thrombin does not occur in plasma. Proc. Natl. Acad. Sci., 2007, 104, 12855-12860.
Matafonov, A.; Leung, P.Y.; Gailani, A.E.; Grach, S.L.; Puy, C.; Cheng, Q.; Sun, M.F.; McCarty, O.J.; Tucker, E.I.; Kataoka, H.; Renné, T.; Morrissey, J.H.; Gruber, A.; Gailani, D. Factor XII inhibition reduces thrombus formation in a primate thrombosis model. Blood, 2014, 123(11), 1739-1746.
[] [PMID: 24408325]
Jansen, P.M.; Pixley, R.A.; Brouwer, M.; de Jong, I.W.; Chang, A.C.; Hack, C.E.; Taylor, F.B., Jr; Colman, R.W. Inhibition of factor XII in septic baboons attenuates the activation of complement and fibrinolytic systems and reduces the release of interleukin-6 and neutrophil elastase. Blood, 1996, 87(6), 2337-2344.
[PMID: 8630396]
Larsson, M.; Rayzman, V.; Nolte, M.W.; Nickel, K.F.; Björkqvist, J.; Jämsä, A.; Hardy, MP.; Fries, M.; Schmidbauer, S.; Hedenqvist, P.; Broomé, M. A factor XIIa inhibitory antibody provides thromboprotection in extracorporeal circulation without increasing bleeding risk. Sci. Transl. Med, 2014, 6(222), 222ra17-222ra17.
Kato, H.; Gomez, E.A.; Fujita, M.; Ishimaru, Y.; Uezato, H.; Mimori, T.; Iwata, H.; Hashiguchi, Y. Ayadualin, a novel RGD peptide with dual antihemostatic activities from the sand fly Lutzomyia ayacuchensis, a vector of Andean-type cutaneous leishmaniasis. Biochimie, 2015, 112, 49-56.
[] [PMID: 25724270]
Yau, J.W.; Liao, P.; Fredenburgh, J.C.; Stafford, A.R.; Revenko, A.S.; Monia, B.P.; Weitz, J.I. Selective depletion of factor XI or factor XII with antisense oligonucleotides attenuates catheter thrombosis in rabbits. Blood, 2014, 123(13), 2102-2107.
[] [PMID: 24501216]
Woodruff, R.S.; Xu, Y.; Layzer, J.; Wu, W.; Ogletree, M.L.; Sullenger, B.A. Inhibiting the intrinsic pathway of coagulation with a factor XII-targeting RNA aptamer. J. Thromb. Haemost., 2013, 11(7), 1364-1373.
[] [PMID: 23692437]
Colman, R.W. Are hemostasis and thrombosis two sides of the same coin? J. Exp. Med., 2006, 203(3), 493-495.
[] [PMID: 16533890]
Gailani, D.; Renné, T. The intrinsic pathway of coagulation: a target for treating thromboembolic disease? J. Thromb. Haemost., 2007, 5(6), 1106-1112.
[] [PMID: 17388803]

Rights & PermissionsPrintExport Cite as

Article Details

Year: 2020
Published on: 10 February, 2020
Page: [243 - 250]
Pages: 8
DOI: 10.2174/0929866526666191026111056
Price: $65

Article Metrics

PDF: 17