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Streptokinase-polyethylene glycol conjugates with increased stability and reduced side effects

Russian Journal of Bioorganic Chemistry volume 40, pages 516–525 (2014)Cite this article

Abstract

Covalent SK-PEG2 and SK-PEG5 conjugates with various degrees of modification of the protein amino groups were obtained by variation of the duration of streptokinase (SK) incubation with activated polyethylene glycol (M 2 and 5 kDa, PEG2 and PEG5); their properties were studied in comparison with the properties of unmodified SK in vitro. SK-PEG2 and SK-PEG5 conjugates with the highest stability in plasma retaining 80% of initial fibrinolytic activity were formed at modification degrees of 54 and 52%, respectively. Interaction of the conjugates with equimolar plasminogen resulted in the formation of plasmin (Pm) activator complexes Pm·SK-PEG2 and Pm·SK-PEG5 with the maximum amidase activity being the same as that of Pm complex with native SK. Catalytic efficiency of plasminogen activation (k Pg/K Pg) was found to be slightly higher (2.84 min−1 μM−1) in case of Pm·SK-PEG2 complex and slightly lower, in case of the Pm·SK-PEG5 complex (1.17 min−1 μM−1), if compared to that of the unmodified complex Pm·SK (2.1 min−1 μM−1). Investigation of lysis kinetics of human plasma clot and depletion of plasminogen and fibrinogen plasma levels under the effect of equal doses of SK in free and conjugated forms demonstrated that SK-PEG2 and SK-PEG5 conjugates possess high thrombolytic activity (89 and 72% to the activity of free SK, respectively) and cause 3.5–4-fold lower side effects than free SK. The SK-PEG2 and SK-PEG5 conjugates with increased stability in plasma and reduced side effects may be used in therapy of thrombotic disorders.

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Abbreviations

pNA:

p-nitroaniline

TNBS:

trinitrobenzenesulfonic acid

PA:

plasminogen activators

Pg:

plasminogen

Pm:

plasmin

PEG:

polyethylene glycol

SK:

streptokinase

SK-PEG:

covalent conjugate of streptokinase and polyethylene glycol

Fg:

fibrinogen

References

  1. Gulba, D.C., Bode, C., Runge, M.S., and Huber, K., Fibrinolysis Proteolysis, 1998, vol. 12,Suppl. 2, pp. 39–58.

    Article  CAS  Google Scholar 

  2. Collen, D. and Lijnen, H.R., Fibrinolysis Proteolysis, 2000, vol. 14, pp. 66–72.

    Article  CAS  Google Scholar 

  3. Bachmann, F., Fibrinolysis, in Thrombosis and Haemostasis, Verstraete, M., Vermylen, J., Lijnen, R., and Arnout, J., Eds., Leuven University Press, 1987, pp. 227–265.

    Google Scholar 

  4. McClintock, D.K. and Bell, P.H., Biochem. Biophys. Res. Commun., 1971, vol. 43, pp. 694–702.

    PubMed  Article  CAS  Google Scholar 

  5. Reddy, K.N.N. and Marcus, G., J. Biol. Chem., 1974, vol. 249, pp. 4851–4857.

    PubMed  CAS  Google Scholar 

  6. Buck, F.F., Hummel, B.C.W., and De Renzo, E.C., J. Biol. Chem., 1968, vol. 243, pp. 3648–3654.

    PubMed  CAS  Google Scholar 

  7. Kunamneni, A., Abdelghani, T.T.A., and Ellaiah, P., J. Thromb. Thrombolysis, 2007, vol. 23, pp. 9–23.

    PubMed  Article  CAS  Google Scholar 

  8. Boxrud, P.D., Fay, W.P., and Bock, P.E., J. Biol. Chem., 2000, vol. 275, pp. 14579–14589.

    PubMed  Article  CAS  Google Scholar 

  9. Ferres, H., Drugs, 1987, vol. 33,Suppl. 3, pp. 33–50.

    PubMed  Article  CAS  Google Scholar 

  10. Bashkov, G.V., Aisina, R.B., Makarov, V.A., Sirotenko, A.V., Popova, G.Yu., Zhitkova, Yu.V., and Kazanskaya, N.F., Farmakol. Toksikol., 1991, vol. 54, pp. 46–50.

    PubMed  CAS  Google Scholar 

  11. Wu, X.C., Ye, R., Duan, Y., and Wong, S.L., Appl. Environ. Microbiol., 1998, vol. 64, pp. 824–829.

    PubMed  CAS  PubMed Central  Google Scholar 

  12. Houng, A., Quen, S., Jean, L.-F., and Reed, G.L., Thromb. Haemost., 1995, vol. 73, pp. 1130–1135.

    Google Scholar 

  13. Chazov, E.I. and Golikov, A.P., Kardiologiia, 1981, vol. 21, pp. 10–14.

    PubMed  CAS  Google Scholar 

  14. Kim, Y.-W. and Kim, D.-C., Yakche Hakhoechi, 1999, vol. 29, pp. 211–216.

    CAS  Google Scholar 

  15. Perkins, W.R., Vaughan, D.E., Plavin, S.R., Daley, W.L., Rauch, J., Lee, L., and Janoff, A.S., Thromb. Haemost., 1997, vol. 77, pp. 1174–1178.

    PubMed  CAS  Google Scholar 

  16. Leach, J.K., O’Rear, E.A., Patterson, E., Miao, Y., and Johnson, A.E., Thromb. Haemost., 2003, vol. 90, pp. 64–70.

    PubMed  CAS  Google Scholar 

  17. Leach, J.K., Patterson, E., and O’Rear, E.A., J. Thromb. Haemost., 2004, vol. 2, pp. 1548–1555.

    PubMed  Article  CAS  Google Scholar 

  18. Mukhametova, L.I., Aisina, R.B., Tyupa, D.V., Medvedeva, A.S., and Gershkovich, K.B., Bioorg. Khim., 2013, vol. 39, pp. 437–444.

    PubMed  CAS  Google Scholar 

  19. Veronese, F.M., Biomaterials, 2001, vol. 22, pp. 405–417.

    PubMed  Article  CAS  Google Scholar 

  20. Webster, R., Didier, E., Harris, P., Siegel, N., Stadler, J., Tilbury, L., and Smith, D., Drug Metab. Dispos., 2007, vol. 35, no. 1, pp. 9–16.

    PubMed  Article  CAS  Google Scholar 

  21. Koide, A., Suzuki, S., and Kobayashi, S., FEBS Lett., 1982, vol. 143, pp. 73–76.

    PubMed  Article  CAS  Google Scholar 

  22. Rajagopalan, S., Gonias, S.L., and Pizzo, S.V., J. Clin. Invest., 1985, vol. 75, pp. 413–419.

    PubMed  Article  CAS  PubMed Central  Google Scholar 

  23. Brucato, F.H. and Pizzo, S.V., Blood, 1990, vol. 76, pp. 73–79.

    PubMed  CAS  Google Scholar 

  24. Greenwald, R.B., Choe, Y.H., McGuire, J., and Conover, C.D., Adv. Drug Deliv. Rev., 2003, vol. 55, pp. 217–250.

    PubMed  Article  CAS  Google Scholar 

  25. Yamaoka, T., Tabata, Y., and Ikada, Y., J. Pharm. Sci., 1994, vol. 83, pp. 601–606.

    PubMed  Article  CAS  Google Scholar 

  26. Beauchamp, C.O., Gonias, S.L., Menapace, D.P., and Pizzo, S.V., Anal. Biochem., 1983, vol. 131, pp. 25–33.

    PubMed  Article  CAS  Google Scholar 

  27. Christensen, L.R., Proc. Soc. Biol. Med., 1941, vol. 46, pp. 674–679.

    Article  CAS  Google Scholar 

  28. Chibber, B.A., Morris, J.P., and Castellino, F.J., Biochemistry, 1985, vol. 24, pp. 3429–3434.

    PubMed  Article  CAS  Google Scholar 

  29. Aisina, R., Mukhametova, L., Gershkovich, K., and Varfolomeyev, S.D., Biochim. Biophys. Acta, 2005, vol. 1725, pp. 370–376.

    PubMed  Article  CAS  Google Scholar 

  30. Aisina, R.B., Mukhametova, L.I., Prisyazhnaya, N.V., Gulin, D.A., Levashov, M.Yu., and Gershkovich, K.B., Russ. J. Bioorg. Chem., 2011, vol. 37, pp. 285–291.

    Article  CAS  Google Scholar 

  31. Wiman, B. and Wallen, P., Eur. J. Biochem., 1973, vol. 36, pp. 25–31.

    PubMed  Article  CAS  Google Scholar 

  32. Babashamsi, M., Razavian, M.H., and Nejadmoghaddam, M.R., AJMB, 2009, vol. 1, pp. 47–51.

    PubMed  CAS  PubMed Central  Google Scholar 

  33. Popova, G.Yu., Eremeev, N.L., Aisina, R.B., and Kazanskaya, N.F., Byull. Eksp. Biol. Med., 1989, no. 5, pp. 561–564.

    Google Scholar 

  34. Astrup, T. and Mullerts, S., Arch. Biochem. Biophys., 1952, vol. 40, pp. 346–351.

    PubMed  Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Chemistry, Moscow State University, Moscow, 119992, Russia

    R. B. Aisina, L. I. Mukhametova, D. V. Tyupa & S. D. Varfolomeev

  2. Emanuel Institute of Biochemical Physics, Russian Academy of Sciences, ul. Kosygina 4, Moscow, 119991, Russia

    K. B. Gershkovich & D. A. Gulin

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  1. R. B. Aisina
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  2. L. I. Mukhametova
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  3. D. V. Tyupa
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  4. K. B. Gershkovich
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  6. S. D. Varfolomeev
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Correspondence to R. B. Aisina.

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Original Russian Text © R.B. Aisina, L.I. Mukhametova, D.V. Tyupa, K.B. Gershkovich, D.A. Gulin, S.D. Varfolomeev, 2014, published in Bioorganicheskaya Khimiya, 2014, Vol. 40, No. 5, pp. 560–570.

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Aisina, R.B., Mukhametova, L.I., Tyupa, D.V. et al. Streptokinase-polyethylene glycol conjugates with increased stability and reduced side effects. Russ J Bioorg Chem 40, 516–525 (2014). https://doi.org/10.1134/S1068162014050021

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  • DOI: https://doi.org/10.1134/S1068162014050021

Keywords

  • streptokinase
  • plasminogen
  • polyethylene glycol conjugate
  • activity
  • stability
  • polyethylene glycol
  • fibrinogen