Biotechnologia Acta


  • Increase font size
  • Default font size
  • Decrease font size
Home Archive 2016 № 3 CALIX[4]ARENE C-145 EFFECTS ON СELLULAR HAEMOSTASIS V. O. Chernyshenko, D. S. Korolova, T. V. Nikolaienko, V. Е. Dosenko, D. O. Pashevin, V. I. Kalchenko, S. O. Cherenok, N. N. Khranovska, L. V. Garmanchuk, E. V. Lugovskoy, S. V. Komisarenko
Print PDF

ISSN 2410-7751 (Print)
ISSN 2410-776X (on-line)

"Biotechnologia Acta" V. 9, No 3, 2016
Р. 37-43, Bibliography 23, English
Universal Decimal Classification: 577.112:612.115


V. O. Chernyshenko1, D. S. Korolova 2, T. V. Nikolaienko2, V. Е. Dosenko 3, D. O. Pashevin 3, V. I. Kalchenko 4, S. O. Cherenok 4, N. N. Khranovska 5, L. V. Garmanchuk2, E. V. Lugovskoy 1, S. V. Komisarenko 1

1 Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kyiv
2ESC «Institute of Biology» Taras Shevchenko National University, Kyiv
3 Bohomolets Institute of Physiology of the National Academy of Sciences of Ukraine, Kyiv
4 Institute of Bioorganic Chemistry and Petrochemistry    of the National Academy of Sciences of Ukraine, Kyiv
5 National Cancer Institute   of Ministry of Health of Ukraine, Kyiv

The aim of the research was to study a potential antithrombotic sodium salt of calix[4]arene-methylene-bis-phosphonic acid (С-145) — on activation and aggregation of platelets in vivo, as well as on proliferation and apoptosis of endothelial cells in the cell culture.

Effects of calix[4]arene С-145 estimated in vitro after addition to the platelet rich plasma, and in vivo after intravenous injection into rabbit bloodstream in equivalent amounts (46 μM). Aggregation of platelets was induced by adenosine diphosphate and detected using aggregometer Solar AP2110. Platelet shape and cytoplasmic granularity were monitored on COULTER EPICS XL Flow Cytometer. The level of tissuetype plasminogen activator — tPA — was estimated using enzyme-linked immunosorbent assay ELISA. Effects of calix[4]arene C-145 on culture of endotelial cells cells was studied using 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide — MTT-test. The population of proliferative pool of cells (G2/M+S) was determined using flow cytometry.

Aggregometry and flow cytometry showed that calix[4]arene C-145 did not activate platelets nor affect their aggregation in vitro. However intravenous injection of calix[4]arene C-145 into the bloodstream of healthy rabbits leads to strong inhibition of platelet aggregation and changes of shape and granularity of most of the platelets after 2 hours of administration. Any additional appearance of endothelial cells activation marker tPA in vivo and any inhibition of calix[4]arene C-145 on proliferation of endothelial cells in cell culture did not observe.

So calix[4]arene C-145 had strong anti-platelet effect in vivo that was not a result of their direct action on platelets or endothelial cells in vitro. This allowed to assume the possibility of calix[4]arene C-145 use as an effective antithrombotic agent.

Key words: calix[4]arene, haemostasis, antithrombotic drugs, fibrin polymerization.

© Palladin Institute of Biochemistry of National Academy of Sciences of Ukraine, 2016

  • References
      1. Lugovskoi E. V., Gritsenko P. G., Koshel T. A., Koliesnik I. O., Cherenok S. O., Kalchenko O. I., Kalchenko V. I., Komisarenko S. V. Calix[4]arene methylenebisphosphonic acids as inhibitors of fibrin polymerization. FEBS J. 2011, V. 278, Р. 1244‒1251.
      2. Chernyshenko V. O., Korolova D. S., Dosenko V. Е., Pashevin D. O., Kalchenko V. I., Pirogova L. V., Chernyshenko T. M., Lugovska O. E., Kravchenko N. А., Makogonenko Y. M., Lugovskoy E. V., Komisarenko S. V. Calix[4]arene C-145 Effects on Plasma Haemostasis. Pharm. Anal. Acta. 2015, V. 6, Р. 406.
      3. Broos K., Feys H. B., De Meyer S. F., Vanhoorelbeke K., Deckmyn H. Platelets at work in primary hemostasis. Blood Rev. 2011, V. 25, Р. 155‒167.
      4. Markland F. S., Swenson S. Fibrolase: Trials and Tribulations. Toxins (Basel). 2010, 2 (4), 793‒808.
      5. Ringwala S. M., Dibattiste P. M., Schneider D. J. Effects on platelet function of a direct acting antagonist of coagulation factor Xa. J. Thromb. Thrombolysis. 2012, 34 (3), 291‒296.
      6. Gryshchuk V., Galagan N. Silica Nanoparticles Effects on Blood Coagulation Proteins and Platelets. Biochem. Res. Int. 2016, V. 2016, Р. 2959414.
      7. Pelagalli A., Scalia G., Pero M. E., Morabito P., Mastellone V., Lombardi P., Vecchio L., Avallone L. Platelet aggregation and flow cytometry analysis of canine platelet rich plasma. Rev. Méd. Vétér. 2008, 10 (159), 481‒484.
      8. Cattaneo M., Cerletti C., Harrison P., Hayward C. P., Kenny D., Nugent D., Nurden P., Rao A. K., Schmaier A. H., Watson S. P., Lussana F., Pugliano M. T., Michelson A. D. Recommendations for the Standardization of Light Transmission Aggregometry: A Consensus of the Working Party from the Platelet Physiology Subcommittee of SSC/ISTH. J. Thromb. Haemost. 2013, V. 11, P. 1183‒1189.
      9. Selected methods for antibody and nucleic acid probes. Cold Spring Harbor Laboratory Press. USA. 1993.
      10. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxic assayas. J. Immunol. Meth. 1983, 65 (1‒2), 55‒63.
      11. Nicoletti I. Migliorati G., Pagliacci M. C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J. Immunol. Meth. 1991, 139 (2), 271‒280.
      12. Verhamme P., Hoylaerts M. F. The pivotal role of the endothelium in haemostasis and thrombosis. Acta Clin. Belg. 2006, 6 (5), 213‒219.
      13. Urano T., Suzuki Y. Accelerated Fibrinolysis and Its Propagation on Vascular Endothelial Cells by Secreted and Retained tPA. J. Biomed. Biotechnol. Volume. 2012, V. 2012, Р. 208108.
      14. Visentin G. P., Liu C. Y. Drug Induced Thrombocytopenia. Hematol. Oncol. Clin. North. Amer. 2007, 2 (4), 685‒686.
      15. Kenney B., Stack G. Drug-Induced Thrombocytopenia. Arch. Pathol. Lab. Med. 2009, V. 133, Р. 309‒314.
      16. Ahmed I., Majeed A., Powell R. Heparin induced thrombocytopenia: diagnosis and management update. Postgrad. Med. J. 2007, V. 83, Р. 575‒582.
      17. Aster R. H. Immune thrombocytopenia caused by glycoprotein IIb/IIIa inhibitors. Chest. 2005, V. 127, Р. 53‒59.
      18. Bougie D. W., Wilker P. R., Wuitschick E. D., Curtis B. R., Malik M., Levine S., Lind R. N., Pereira J., Aster R. H. Acute thrombocytopenia after treatment with tirofiban or eptifibatide is associated with antibodies specific for ligand-occupied GPIIb/IIIa. Blood. 2002, 100 (6), 2071‒2076.
      19. Wazny L. D., Ariano R. E. Evaluation and management of drug-induced thrombocytopenia in the acutely ill patient. Pharmacotherapy. 2000, 20 (3), 292‒307.
      20. Majhail N. S., Lichtin A. E. What is the best way to determine if thrombocytopenia in a patient on multiple medications is drug-induced? Cleve Clin. J. Med. 2002, 69 (3), 259‒262.
      21. Sinzinger H., Fitscha P., Peskar B. A. Platelet half-life, plasma thromboxane B2 and circulating endothelial-cells in peripheral vascular disease. Angiology. 1986, 37 (2), 112‒118.
      22. Lindblad B., Wright S. W., Burkel W. E., Wakefield T. W., Graham L. M., Sell R., Stanley J. C. Endothelial cell proliferation in vitro after incubation with anticoagulant and antiplatelet drugs. Artery. 1988, 16 (1), 15‒24.
      23. Mintzer D. M., Billet S. N., Chmielewski L. Drug-Induced Hematologic Syndromes. Adv. Hematol. 2009, V. 2009, Р. 495863.


Additional menu

Site search

Site navigation

Home Archive 2016 № 3 CALIX[4]ARENE C-145 EFFECTS ON СELLULAR HAEMOSTASIS V. O. Chernyshenko, D. S. Korolova, T. V. Nikolaienko, V. Е. Dosenko, D. O. Pashevin, V. I. Kalchenko, S. O. Cherenok, N. N. Khranovska, L. V. Garmanchuk, E. V. Lugovskoy, S. V. Komisarenko

Invitation to cooperation

Dear colleagues, we invite you to publish your articles in our journal.
© Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, 2008.
All rights are reserved. Complete or partial reprint of the journal is possible only with the written permission of the publisher.
for information: