Fibrin-specific thrombolytic agents

The mammalian fibrinolytic system comprises a proenzyme, plasminogen, which can be converted to the active enzyme plasmin, which will degrade fibrin. Plasminogen activation is mediated by plasminogen activators which are classified as either tissue-type plasminogen activator (t-PA) or urokinase-type plasminogen activator (u-PA). t-PA and single-chain u-PA (scu-PA) induce clot-specific thrombolysis, however via entirely different mechanisms. t-PA is relatively inactive in the absence of fibrin, but fibrin strikingly enhances the activation rate of plasminogen by t-PA. This is explained by an increased activity of fibrin-bound t-PA for plasminogen and not by alteration of the catalytic efficiency of the enzyme. scu-PA has a high affinity for plasminogen but, however, does not activate plasminogen in plasma in the absence of a fibrin clot, due to competitive inhibition. Fibrin-specific thrombolysis appears to be due to the fact that fibrin reverses the competitive inhibition, but this does not seem to occur via specific binding of scu-PA to fibrin. The thrombolytic efficacy and fibrin-specificity of natural and recombinant t-PA has been demonstrated in animal models of pulmonary embolism, venous thrombosis and coronary artery thrombosis. In all these studies thrombolysis and relative fibrinogen sparing effect of t-PA was recently confirmed in several multicenter clinical trials in patients with acute myocardial infarction. Specific thrombolysis by scu-PA has also been demonstrated in animal models of pulmonary embolism, venous thrombosis and coronary artery thrombosis.     

New thrombolytic agents

Tissue Plasminogen Activator (t-PA), Single Chain Urokinase Plasminogen Activator or pro-Urokinase (scu-PA or pro-UK) and acyl enzymes are new thrombolytic agents, characterized by a high fibrin affinity, so that they provoke only mild systemic fibrinolytic effect. Their infusion would allie good thrombolytic activity and reduced hemorragic risks, usually related to fibrinogen and others coagulation factors degradation t-PA and scu-PA are natural, physiological substances, obtained by recombinant DNA technology. t-PA infusion in acute myocardial infusion (AMI) has been shown to be at least as efficient than intracoronary Streptokinase (SK) administration, but fibrinogenolysis was much lower as compared to SK. In vitro studies have shown that scu-PA was an efficient thrombolytic agent and has a relative fibrin specificity, at least as similar to t-PA, but much superior to classical Urokinase. The acyl-enzyme APSAC or Eminase is a SK-plasminogen complex in which the proteolytic site has been inactivated with an anisoic-acid. This acyl enzyme has a longer half-life than t-PA and scu-PA, and can be injected as a bolus. Its administration in AMI have shown that APSAC is as effective as SK, but can also provoke severe fibrinogenolysis. These 3 agents seem to have similar thrombolytic activities on coronary thrombi. However, further studies are required to evaluate the bleeding incidence and coronary reocclusion rates associated with their utilisation.     

Prothrombin activation by a metalloprotease from Staphylococcus aureus.

Formation of thrombin during incubation of purified bovine prothrombin with purified staphylococcal metalloprotease has been investigated. Thrombin activity was estimated by examination of clotting time and by digestion of a synthetic substrate, Chromozym TH. The metalloprotease caused direct activation of prothrombin which was inhibited by the addition of ethylenediaminetetraacetic acid. Metalloprotease produced by some strains of Staphylococcus aureus may simulate staphylocoagulase activity.     

Prothrombin activation in eastern tiger snake bite

AIMS: To perform ex vivo studies in eastern tiger snake envenomation which define the haemostatic events associated with prothrombin activation. METHOD: Serial studies were performed on plasma from six individuals with evidence of eastern tiger snake envenomation. These analyses were particularly directed to fibrinogen levels, F1 + 2, TAT and evidence of fibrinolysis. RESULTS: There was a substantial rise in F1 + 2 and thrombin-antithrombin (TAT) complexes in all cases, even with minimal evenomation. In some cases the molar ratio of F1 + 2 and TAT was reduced from the relationship normally seen in vitro and ex vivo in clinical thrombosis. There was a dramatic fall in factor V and VIII levels which occurred 4-6 hours before the decline in prothrombin and AT3. This related in time to a fall in alpha2AP and plasminogen. Protein C levels also declined dramatically but many hours after presentation. CONCLUSIONS: F1 + 2 and TAT are sensitive markers of tiger snake evenomation. In some patients with massive prothrombin activation, the common mechanism for TAT clearance may be altered or overwhelmed. Conversely, the renal clearance of the smaller F1 + 2 may be enhanced. In the absence of thrombocytopaenia, which is a very sensitive marker of DIC, the fall in labile factors with tiger snake envenomation is significantly contributed to by proteolytic digestion of clotting factors.     

Intraperitoneal thrombolytic agents in relapsing or persistent peritonitis of patients on continuous ambulatory peritoneal dialysis

Urokinase or streptokinase was instilled intraperitoneally as an adjunct to the antibiotic therapy in 16 episodes of relapsing or persistent peritonitis in CAPD patients. In eight patients the combination of antibiotics and intraperitoneal thrombolytic agents resulted in clearing of the infection with no recurrences. The treatment failed in eight other patients, who had their peritoneal catheters removed. Six of the last eight patients had either abdominal wall abscesses or persistence of the bacteria on the wall of the peritoneal catheter. Elevated post-intraperitoneal instillation peritoneal fluid neutrophil counts and positive post-instillation peritoneal fluid cultures predicted failure of the intraperitoneal instillation of thrombolytic agents in most instances. Intraperitoneal instillation of urokinase or streptokinase may help cure approximately 50% of the episodes of relapsing for persistent peritonitis. Post-instillation peritoneal fluid cell counts and cultures should be monitored. Radiologic investigation for abdominal wall or intraabdominal abscesses is indicated if intraperitoneal instillation of urokinase or streptokinase fails to eradicate peritonitis.     

Prothrombin time and its standardization

This review regarding prothrombin time and its standardization is described around some recent topics as the followings. 1. History of standardization for prothrombin time and revised WHO guideline for thromboplastin; A short history of standardization is summarized to understand a scheme of International Normalized Ratio (INR) based on International Sensitivity Index (ISI) that is calibrated by International Reference Preparation (IRP) for thromboplastin, and some key points in revised WHO guideline for thromboplastin and plasma used to control oral anticoagulant therapy are interpreted for research and practical use. 2. Point-of-care prothrombin time monitoring; A portable device to measure prothrombin time with whole blood sample, such as CoagChek (Roche), contributes to self-management by patients required long-term oral anticoagulation. Some investigators reported clinical agreement to use this monitoring system and improvement of patient's QOL and cost-effectiveness in overseas. 3. New types of thromboplastins; Two new types of thromboplastins have been available since the last year in Japan. One is a human plain thromboplastin, Simplastin HTF (Bioméreux) from extract of cultured human lung cancer cell, and another is IL test PT-Fibrinogen Recombinant (Iatron) from recombinant rabbit tissue factor relipidated in a synthetic phospholipid blend. For control of oral anticoagulation, good performance are expected in either thromboplastins because of their sufficient low ISI values. 4. INR methodology for other diseases; INR/ISI system is designed as a standardized methodology for control of oral anticoagulation. Prothrombin time, however is utilized as a global coagulation test for diagnosis or criteria of other disorders, such as congenital coagulation factor deficient, severe liver dysfunction and disseminated intravascular coagulation. Previous our study indicated that discrepancy of sensitivities to plasma absorbed multiple coagulation factors and plasma from patients under oral anticoagulation was revealed in rabbit brain thromboplastins, but not in human origins. Discrepancy of sensitivities observed in rabbit thromboplastins was emphasized in convert to INR values. These results suggested that the use of human thromboplastin of which ISI is close to 1.0 leads possibility for introducing INR methodology to evaluate PT of other disorders.