An inhibitor of activated thrombin-activatable fibrinolysis inhibitor potentiates tissue-type plasminogen activator-induced thrombolysis in a rabbit jugular vein thrombolysis model.
When activated in vitro, thrombin-activatable fibrinolysis inhibitor (TAFI) slows clot lysis by cleaving the C-terminal lysine and arginine residues from partially degraded fibrin. An inhibitor of carboxypeptidase isolated from potato (CPI) reverses prolongation of clot lysis by inhibiting activated TAFI. We investigated in vivo effect of TAFI inhibition on tissue-type plasminogen activator (t-PA)-induced clot lysis using CPI in a rabbit jugular vein thrombolysis model. It was found necessary to further purify the CPI preparations from commercial sources by HPLC chromatography to remove endotoxin and anti-plasmin activity that would affect the endogenous fibrinolytic system. The effect of intravenous administration of the purified CPI with t-PA was determined by measuring thrombus weight at the end of 90 minutes in six groups of animals. In the control group receiving saline, the median thrombus weight was 116 mg. In the group that received CPI only (0.5 mg/kg bolus injection followed by 0.3 mg/kg/h infusion), the median thrombus weight was 121 mg. In the group that received t-PA at a dose of 10 microg/kg bolus followed by 67 microg/kg/h infusion, the median thrombus weight decreased to 86 mg. When CPI was coadministered with the same regimen of t-PA, the median value further decreased to 58 mg. When animals were given three times higher the dose of t-PA (30 microg/kg bolus followed by 200 microg/kg/h infusion) in the absence or presence of CPI, median thrombus weights were 56 mg and 0 mg, respectively. Our results demonstrate that systemic coadministration of the purified CPI improves clot lysis induced by t-PA.
Nagashima
M
M
Department of Cardiovascular Research, Berlex Biosciences, Richmond, CA, USA. Mariko_Nagashima@berlex.com
Werner
M
M
Wang
M
M
Zhao
L
L
Light
D R
DR
Pagila
R
R
Morser
J
J
Verhallen
P
P
D000818:Animals; D025901:Carboxypeptidase B2; D002268:Carboxypeptidases / Q000037:antagonists & inhibitors* / Q000502:physiology; D002851:Chromatography, High Pressure Liquid; D004353:Drug Evaluation, Preclinical; D004357:Drug Synergism; D004730:Endothelium, Vascular / Q000503:physiopathology; D004789:Enzyme Activation / Q000187:drug effects; D005337:Fibrin / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism; D005342:Fibrinolysis / Q000187:drug effects*; D007262:Infusions, Intravenous; D010940:Plant Proteins / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology / Q000627:therapeutic use*; D010958:Plasminogen / Q000378:metabolism; D011480:Protease Inhibitors; D011817:Rabbits; D011198:Solanum tuberosum / Q000201:enzymology; D013917:Thrombin / Q000502:physiology*; D015912:Thrombolytic Therapy; D010959:Tissue Plasminogen Activator / Q000494:pharmacology / Q000627:therapeutic use*; D020246:Venous Thrombosis / Q000188:drug therapy*; D000979:alpha-2-Antiplasmin / Q000378:metabolism
D010940:Plant Proteins; D011480:Protease Inhibitors; D000979:alpha-2-Antiplasmin; D005337:Fibrin; D005340:Fibrinogen; D010958:Plasminogen; D002268:Carboxypeptidases; D025901:Carboxypeptidase B2; D013917:Thrombin; D010959:Tissue Plasminogen Activator