Effects of the synthetic thrombin inhibitor argatroban on fibrin- or clot-incorporated thrombin: comparison with heparin and recombinant Hirudin.
The inhibitory effects of argatroban on clot- or fibrin-bound human thrombin were studied using the thrombin-specific chromogenic substrate S2238 (200 microM). These effects were compared to those of recombinant hirudin (rHV2 Lys 47) and the heparin/antithrombin III complex. Argatroban concentration-dependently inhibited the cleavage of S2238 by a thrombin solution, which had been titrated to give the same change in OD405 nm as fibrin-bound thrombin, with an IC50 of 1.1 microM with 90% inhibition at 8 microM. rHV2 Lys 47 and heparin had IC50 values of 1.2 nM and 0.003 U/ml respectively under these conditions. However, when the compounds were tested against fibrin-bound thrombin, argatroban had an IC50 of 2.8 microM with 65% inhibition at 8 microM, whereas rHV2 Lys 47 had an IC50 of 23 nM (with only 56% inhibition at 200 nM), and heparin had an IC50 of 0.5 +/- 0.38 U/ml (with only 58% inhibition at 5 U/ml); i. e. the two compounds were 19 and 168 times less active against fibrin-bound thrombin than against thrombin in solution. The differences between the inhibitory effects of the compounds against thrombin bound to a plasma clot were even more striking in that the IC50 of argatroban was increased from 1.1 (vs. thrombin in solution) to 2.7 microM, while, although rHV2 Lys 47 and heparin had IC50 values of 2.8 nM and 0.004 U/ml against thrombin in solution, they had little (32% inhibition by 4 microM rHV2 Lys 47) or no effect (even at 5.0 U/ml heparin) against the amidolytic activity of a plasma clot.(ABSTRACT TRUNCATED AT 250 WORDS)
Berry
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Pre-Clinical Research Department, Synthélabo Recherche (L.E.R.S.), Bagneux, France.
Girardot
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Lecoffre
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Lunven
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D000991:Antithrombins / Q000494:pharmacology*; D001120:Arginine / Q000031:analogs & derivatives; D001777:Blood Coagulation; D004151:Dipeptides / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005337:Fibrin; D006493:Heparin / Q000494:pharmacology*; D006629:Hirudins / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D010875:Pipecolic Acids / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000494:pharmacology; D013449:Sulfonamides; D013917:Thrombin / Q000037:antagonists & inhibitors*
D000991:Antithrombins; D004151:Dipeptides; D006629:Hirudins; D010875:Pipecolic Acids; D011994:Recombinant Proteins; D013449:Sulfonamides; C064761:hirudin, Lys(47)-; C021129:S 2238; D005337:Fibrin; D006493:Heparin; D001120:Arginine; D013917:Thrombin; C031942:argatroban