350 rub
Journal Technologies of Living Systems №8 for 2009 г.
Article in number:
FUNCTIONAL STUDY OF THE NEW SYNTHETIC THROMBIN INHIBITOR HC-019S-IOC IN VITRO AND ITS TRIAL IN VIVO AS A PART OF A PLASMA SUBSTITUTING SOLUTION IN MODEL OF HEMODILUTION IN RATS
Keywords: blood coagulation synthetic thrombin inhibitor animal hemodilution model rats endogenous thrombin potential spatial clot growth rate
Authors:
E.I. Sinauridze, S.S. Surov, I.V. Gribkova, A.S. Gorbatenko, M.Y. Monakov, F.I. Ataullakhanov
Abstract:
We have shown before that moderate plasma dilution in vitro (up to 2-2.5 folds) with different ar-tificial plasma substituting solutions (PSS) produced hypercoagulant changes in coagulation sys-tem: the endogenous thrombin potential (ETP) and spatial clot growth rate (Vclot) were increased. Evidently it is consequence of plasma coagulation inhibitors - dilution, first of all antithrombin III. The supplementation of PSS with thrombin inhibitors really corrected these disturbances in vitro. Starting from these results we decided to create a new PSS, supplemented with thrombin inhibitor. Infusion of such solution should not produce hemodilutional hypercoagulation. In this study one of our newly designed low molecular weight synthetic thrombin inhibitors (HC-019s-IOC) was investigated in details: its inhibitory activity towards thrombin in buffer solu-tion, and its anticoagulant effectiveness in blood plasma. It was shown that this inhibitor-s effec-tiveness exceeded the one for argatroban - the only low molecular weight synthetic thrombin inhi-bitor, which is licenced for intravenous administration during thrombotic situation correction in some countries today. Inhibition constant of the new inhibitor (for thrombin in buffer solution) and its IC50 for ETP and Vclot decrease are 0.8 nM, 0.25 µM, and 12 µM, respectively. The constant for thrombin inhibition (in buffer solution), and IC50 for ETP decrease in plasma for argatroban are 39 nM and 0.65 µM, respectively. Moreover, the acute toxicity of this new inhibitor was studied (mouse, intraperitoneal administration) and was found to be 419±33.0 mg/kg. This value is a little worse than toxicity for argatroban (475 mg/kg, mouse, intraperitoneal administration), but it is quite acceptable taking into account the better effectiveness of our inhibitor. Examination of the HC-019s-IOC stability after autoclaving and during long-term storage in saline have shown that this compound did not lose inhibitory activity during autoclaving and long-term storage (>1.5 years) at +40C and at room temperature. So, our new thrombin inhibitor is very promising for usage in PSS. In the second part of the study the model of moderate (23.0±4.5%) hemodilition in vivo in rats was developed. It have been shown using this model that hemodilution by saline really produced hypercoagulation state in animals. It agrees well with our previous in vitro results. The supple-mentation of the saline with 2 µM of the new thrombin inhibitor HC-019s-IOC prevented hyper-coagulation. So, we suppose that new PSS supplemented with thrombin inhibitor may be very prospective for prevention of hypercoagulation produced by hemodilution. New investigations are required to confirm a possibility of such strategy in animal models and in clinic.
Pages: 67-77
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