Inhibition of endothelial cell-mediated generation of activated protein C by direct and antithrombin-dependent thrombin inhibitors.
The present study investigated the effect of the thrombin inhibitors antithrombin (AT) (with and without unfractionated heparin or low molecular weight heparin), hirudin, inogatran and melagatran on thrombin-thrombomodulin-mediated generation of activated protein C (APC), in solution and on endothelial cells. Sequential incubation with thrombin, thrombin inhibitors and protein C was followed by measurement of APC by an amidolytic assay. The approximate concentrations resulting in 50% inhibition of endothelial cell-mediated APC generation for AT, AT-unfractionated heparin, AT-low molecular weight heparin, hirudin, melagatran and inogatran were 200, 4, 9, 1, 8 and 60 nmol/l, respectively. The normal plasma level of AT is 2800 nmol/l and relevant therapeutic concentrations from clinical trials are 200 nmol/l for hirudin, 500 nmol/l for melagatran and 1000 nmol/l for inogatran. The present study indicates that clinically relevant concentrations of the tested thrombin inhibitors interfere with endothelial-mediated APC generation, which may offer an explanation for the lack of a dose-response effect in clinical trials with thrombin inhibitors.
Linder
Rikard
R
Department of Cardiology, Karolinska Hospital, Stockholm, Sweden and Department of Surgical Sciences, Karolinska Institutet, Stockholm, Sweden. rikard.linder@medks.ki.se
Frebelius
Siw
S
Jansson
Katarina
K
Swedenborg
Jesper
J
D000991:Antithrombins / Q000494:pharmacology*; D001384:Azetidines; D001596:Benzylamines; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005434:Flow Cytometry; D005998:Glycine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D006629:Hirudins / Q000494:pharmacology; D006801:Humans; D010880:Piperidines / Q000494:pharmacology; D011486:Protein C / Q000378:metabolism*; D013917:Thrombin / Q000037:antagonists & inhibitors* / Q000502:physiology; D018180:Thrombomodulin / Q000502:physiology
D000991:Antithrombins; D001384:Azetidines; D001596:Benzylamines; D006629:Hirudins; D010880:Piperidines; D011486:Protein C; D018180:Thrombomodulin; C109573:melagatran; C099871:inogatran; D006493:Heparin; D013917:Thrombin; D005998:Glycine