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Cationic chitosan derivative for heparin reversal - in vitro and in vivo studies in rats

Kamil K. Kamiński 1Barbara Lorkowska 2Justyna Ciejka 1Karolina Zazakowny 1Krzysztof Szczubiałka 1Maria Nowakowska 1Ryszard Korbut 2

1. Jagiellonian University, Faculty of Chemistry, Ingardena 3, Kraków 30-060, Poland
2. Jagiellonian University Medical College Chair of Pharmacology (UJCM), Grzegórzecka 16, Kraków 31-531, Poland


Heparin is a widely used anticoagulant agent, which unfortunately may show some side effects, including hemorrhages, osteoporosis and thrombocytopenia. Moreover, the therapeutic dose of heparin varies significantly depending on the patient and therefore cases of overdose are frequent. At present, if necessary, heparin is neutralized using protamine sulfate. That drug, an exogenous protein, can cause an allergic reaction after administration. So, there is a need for an alternative system which would efficiently neutralize heparin, while not inducing allergic reaction.

Recently, we have developed cationic chitosan derivatives in reaction with glycidyltrimethylammonium chloride [1] which have been shown to bind heparin efficiently in buffer solution and in human blood plasma.

In a current presentation we would like to show the preliminary results of in vitro and in vivo studies of chitosan derivative (ChGl2) as a heparin antidote using Wistar rat model. Effect of GhGl2 on heparin activity in animal blood was determined using aPTT (activated Partial Thromboplastin Time) tests. Our study revealed that in rat blood in vitro ChGl2 can neutralize anticoagulant activity of heparin as revealed by significant aPTT reduction. The dose of ChGl2 required for heparin neutralization was determined. Also, comparative in vitro studies on protamine interaction with heparin in animal blood were carried out.

During in vivo experiments, defined dose of heparin was injected into femoral vein of the animal. Than, a fixed dose (predetermined in in vitro tests) of ChGl2 was intravenously administrated to the animal. Blood samples were collected from the carotid artery at regular intervals and aPTT values in obtained samples were determined. The decrease aPTT levels followed the pattern observed in control experiments in which the protamine sulfate was used as an heparin reversal agent. The studies were completed by the ChGl2 toxicity tests.

Based on the experiments described above it was concluded that chitosan derivative (ChGl2) functions as an efficient heparin antidote in rats. The dose of ChGl2 required to bring down the heparin level in rat model is comparable to that of protamine. Further studies on larger population of animals are necessary to confirm the above conclusions and to bring the chitosan derivatives to the clinical studies.


1.  K. Kamiński, K. Zazakowny, K. Szczubialka, M. Nowakowska, pH-Sensitive Genipin-Cross-Linked Chitosan Microspheres For Heparin Removal Biomacromolecules 2008, 9, 3127–3132

Acknowledgements.  Project operated within the Foundation for Polish Science Team Programme (PolyMed, TEAM/2008-2/6) and Ventures Programme co-financed by the EU European Regional Development Fund. Project partially financed by a grant from Polish Ministry of Science and Higher education No. N N204 151336.


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Presentation: Oral at VII Multidyscyplinarna Konferencja Nauki o Leku, by Kamil K. Kamiński
See On-line Journal of VII Multidyscyplinarna Konferencja Nauki o Leku

Submitted: 2010-02-24 21:40
Revised:   2010-03-04 13:53