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D Collen

Publications and source records attributed to D Collen.

At least 433 records · Page 24Linked to original sources
PubMed1988

Characterization of a chimeric plasminogen activator consisting of amino acids 1 to 274 of tissue-type plasminogen activator and amino acids 138 to 411 of single-chain urokinase-type plasminogen activator.

A chimeric plasminogen activator (t-PA/scu-PA-s), consisting of amino acids 1-263 of tissue-type plasminogen activator (t-PA) and 144-411 of single-chain urokinase-type plasminogen activator (scu-PA), was previously shown to maintain the enzymatic properties of scu-PA but to have only partially acquired the fibrin affinity of t-PA, possibly as a result of steric interaction between the functional domains of t-PA and scu-PA (Nelles, L., Lijnen, H. R., Collen, D., and Holmes, W.E. (1987) J. Biol. Chem. 262, 10855-10862). Therefore, we now have constructed an extended chimeric t-PA/scu-PA protein, consisting of amino acids 1-274 of t-PA and 138-411 of scu-PA, which thus has an additional sequence of 17 residues in the region joining the two proteins. The highly purified extended chimeric protein (t-PA/scu-PA-e) was found to have similar specific activity on fibrin film (65,000 IU/mg), kinetic constants for the activation of plasminogen (Km = 1 microM, k2 = 0.0026 s-1), fibrin affinity (50% binding at a fibrin concentration of 3.3 g/liter), and fibrin specificity of clot lysis in a plasma environment (50% lysis in 2 h with 8 nM of the chimer) as the previously characterized chimeric protein (t-PA/scu-PA-s). Thus, unexpectedly, the fibrin affinity of t-PA is also only partially expressed in this extended chimeric protein. Therefore, the NH2-terminal chains (A-chains) of the plasmin-generated two-chain derivatives t-PA/tcu-PA-e, t-PA/tcu-PA-s, and of t-PA were isolated. These A-chain structures of the chimers were found to have lost most of their fibrin affinity, whereas the fibrin affinity of the A-chain of native t-PA was maintained. Differential reactivity of the A-chain structures of both chimeric molecules with monoclonal antibodies directed against the A-chain of t-PA suggested that they were conformationally altered. Sequential fibrin binding experiments with t-PA/scu-PA-e and t-PA/scu-PA-s yielded 45 +/- 8 (n = 11) and 43 +/- 5% (n = 8), respectively, binding in the first cycle and 44 +/- 7 (n = 11) and 27 +/- 10% (n = 8), respectively, binding in the second cycle. This suggests that the low affinity of the chimeric molecules for fibrin is not due to the occurrence of subpopulations of molecules with different fibrin affinity but, instead, to a uniformly decreased fibrin affinity in all molecules.

Amino Acid Sequence↗
PubMed1988

Structural and functional characterization of mutants of recombinant single-chain urokinase-type plasminogen activator obtained by site-specific mutagenesis of Lys158, Ile159 and Ile160.

Single-chain urokinase-type plasminogen activator (scu-PA) is converted to urokinase by hydrolysis of the Lys158-Ile159 peptide bond. Site-directed mutagenesis of Lys158 to Gly or Glu yields plasmin-resistant mutants with a 10-20-fold reduced catalytic efficiency for the activation of plasminogen [Nelles et al. (1987) J. Biol. Chem. 262, 5682-5689]. In the present study, we have further evaluated the enzymatic properties of derivatives of recombinant scu-PA (rscu-PA), produced by site-directed mutagenesis of Lys158, Ile159 or Ile160, in order to obtain additional information on the structure/function relations underlying the enzymatic properties of the single- and two-chain u-PA moieties. [Arg158]rscu-PA (rscu-PA with Lys158 substituted with Arg) appeared to be indistinguishable from wild-type rscu-PA with respect to plasminogen-activating potential (catalytic efficiency k2/Km = 0.21 mM-1 s-1 versus 0.64 mM-1 s-1), conversion to active two-chain urokinase by plasmin (k2/Km = 0.13 microM-1 s-1 versus 0.28 microM-1 s-1), as well as its specific activity (48,000 IU/mg as compared to 60,000 IU/mg) and its fibrinolytic potential in a plasma medium (50% lysis in 2 h with 2.8 micrograms/ml versus 2.1 micrograms/ml). [Pro159]rscu-PA (Ile159 substituted with Pro) and [Gly159]rscu-PA (Ile159 converted to Gly) are virtually inactive towards plasminogen (k2/Km less than 0.004 mM-1 s-1). They are however converted to inactive two-chain derivatives by plasmin following cleavage of the Arg156-Phe157 peptide bond in [Pro159]rscu-PA and of the Lys158-Gly159 peptide bond in [Gly159]rscu-PA. [Gly158,Lys160]rscu-PA (with Lys158 converted to Gly and Ile160 to Lys) has a low catalytic efficiency towards plasminogen both as a single-chain form (k2/Km = 0.012 mM-1 s-1) and as the two-chain derivative (k2/Km = 0.13 mM-1 s-1) generated by cleavage of both the Arg156-Phe157 and/or the Lys160-Gly161 peptide bonds by plasmin. These findings suggest that the enzymatic properties of rscu-PA are critically dependent on the amino acids in position 158 (requirement for Arg or Lys) and position 159 (requirement for Ile). Conversion of the basic amino acid in position 158 results in a 10-20-fold reduction of the catalytic efficiency of the single-chain molecule but yields a fully active two-chain derivative. The presence of Ile in position 159 is not only a primary determinant for the activity of the two-chain derivative, but also of the single-chain precursor. Cleavage of the Arg156-Phe157 or the Lys160-Gly161 peptide bonds by plasmin yields inactive two-chain derivatives.

Animals↗
PubMed1988

Inhibition in purified systems and in human plasma of chimaeric plasminogen activators consisting of the NH2-terminal region of tissue-type plasminogen activator and the COOH-terminal region of urokinase-type plasminogen activator.

Recombinant chimaeric molecules between tissue-type plasminogen activator (t-PA) and single chain urokinase-type plasminogen activator (scu-PA) or two chain urokinase-type plasminogen activator (tcu-PA) have intact enzymatic properties of scu-PA or tcu-PA towards natural and synthetic substrates (Nelles et al., J. Biol Chem 1987; 262: 10855-10862). In the present study, we have compared the reactivity with inhibitors of both the single chain and two chain variants of recombinant u-PA and two recombinant chimaeric molecules between t-PA and scu-PA (t-PA/u-PA-s: amino acids 1-263 of t-PA and 144-411 of u-PA; t-PA/u-PA-e: amino acids 1-274 of t-PA and 138-411 of u-PA). Incubation with human plasma in the absence of a fibrin clot for 3 h at 37 degrees C at equipotent concentrations (50% clot lysis in 2 h), resulted in significant fibrinogen breakdown (to about 40% of the normal value) for all two chain molecules, but not for their single chain counterparts. Preincubation of the plasminogen activators with plasma for 3 h at 37 degrees C, resulted in complete inhibition of the fibrinolytic potency of the two chain molecules but did not alter the potency of the single chain molecules. Inhibition of the two chain molecules occurred with a t1/2 of approximately 45 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Antifibrinolytic Agents↗
PubMed1988

Purification and characterization of a plasminogen activator inhibitor 1 binding protein from human plasma. Identification as a multimeric form of S protein (vitronectin).

A binding protein for plasminogen activator inhibitor 1 (PAI-1-BP) was isolated from human plasma by a four-step procedure. 1) The 7 S globulin fraction of plasma was isolated by gel filtration on Sephacryl S-300. 2) Human endothelial cell-type plasminogen activator inhibitor (PAI-1), pretreated with 12 M urea, was added to this fraction (22 micrograms of PAI-1/ml of plasma), and a PAI-1 antigen peak with apparent mass 450 kDa (representing 65% of PAI-1 antigen and 85% of PAI activity) was isolated by gel filtration of this mixture. 3) The PAI-1.PAI-1-BP complex was further purified by immunoadsorption on an immobilized murine monoclonal antibody directed against PAI-1 (MA-7D4) and by elution with 4 M KSCN. 4) The complex was then dissociated by addition of excess human tissue-type plasminogen activator (t-PA), and t-PA and PAI-1 antigen (t-PA.PAI-1 complexes and free t-PA and PAI-1) were removed by immunoadsorption on monoclonal antibodies directed against t-PA (MA-62E8) and against PAI-1 (MA-7D4 and MA-12A4). Sodium dodecyl sulfate-gel electrophoresis of the purified material under nonreducing conditions revealed two bands with apparent mass approximately equal to 150 kDa and two bands with mass 74 and 68 kDa. Reduced sodium dodecyl sulfate-gel electrophoresis displayed two main bands with apparent masses of 73 and 64 kDa. The PAI-1-BP reacts with urea-treated, but not with inactive PAI-1. t-PA dissociates the complex between PAI-1 and PAI-1-BP. PAI-1 in complex with PAI-1-BP is 2-3-fold more stable at 37 degrees C than purified PAI-1, suggesting that PAI-1-BP may stabilize PAI-1 in blood. The concentration of PAI-1-BP in plasma determined by titration with PAI-1 is approximately 130 mg/liter. The isolated PAI-1-BP was shown to be identical to S protein (vitronectin) both by cross-reactivity with monospecific rabbit antisera and by NH2-terminal amino acid sequence analysis. The gel filtration behavior, mobility on sodium dodecyl sulfate-gel electrophoresis, and concentration in plasma suggest that PAI-1-BP is a multimer (presumably a dimer) of S protein accounting for approximately 35% of the S protein in plasma.

Amino Acids↗
PubMed1988

Functional effects of asparagine-linked oligosaccharide on natural and variant human tissue-type plasminogen activator.

The role of Asn-linked oligosaccharide in the functional properties of both human tissue-type plasminogen activator (t-PA) and a genetic variant of t-PA was studied. Nonglycosylated and glycosylated wild-type t-PA were produced in mammalian cells which express recombinant t-PA. These proteins were compared in fibrin binding and 125I-labeled fibrin clot lysis assays, using purified components. The nonglycosylated form showed higher fibrin binding, as well as higher fibrinolytic potency than the glycosylated form. Subsequently, prevention of glycosylation of a t-PA variant which lacked the finger and epidermal growth factor domains (delta FE), was carried out in an attempt to enhance its fibrinolytic activity. Glycosylation was prevented by changing Asn to Gln; at Asn-117 to produce delta FE1X t-PA, and at Asn-117, -184, and -448 to produce delta FE3X t-PA. All variants were similar to wild-type t-PA in their catalytic dependence on fibrinogen fragments, fibrinolytic activity in fibrin autography analysis, and plasminogen activator activity. In a clot lysis assay, using citrated human plasma, the fibrinolytic potency of the variants were comparable to that of wild-type t-PA at activator concentrations of 17-51 nM (approximately 1-3 micrograms/ml). At 0.5-5.1 nM (approximately 0.03-0.3 micrograms/ml), however, the variant proteins had lower fibrinolytic potency than wild-type t-PA. Fifty percent lysis in 1.5 h for wild-type, delta FE, delta FE1X, and delta FE3X t-PA, required 2.5, 10, 7.5, and 5.5 nM t-PA, respectively. The fibrinogenolytic activity in human plasma was measured for wild-type, delta FE, delta FE1X, and delta FE3X t-PA, and showed significant fibrinogen depletion after 3 h of incubation at 51 nM, decreasing to 11, 11, 50, and 72% of basal levels, respectively. These data indicate that partial or total nonglycosylated t-PA variants have a higher fibrinolytic versus fibrinogenolytic ratio than their fully glycosylated counterparts.

Asparagine↗
PubMed1988

Purification and characterization of natural and recombinant human plasminogen activator inhibitor-1 (PAI-1).

Human plasminogen activator inhibitor-1 (PAI-1) was purified from the conditioned medium of endotoxin-stimulated umbilical vein endothelial cell cultures by combinations of zinc-chelate-Sepharose chromatography, gel filtration on Sephacryl S-300 and immunoadsorption on an insolubilized murine monoclonal antibody (MA-7D4). The final product was obtained with a recovery of approximately 20% from conditioned medium containing about 3 micrograms/ml PAI-1. The yield of PAI-1 was 15-100 micrograms/umbilical cord, depending on the culture and harvest conditions. SDS gel electrophoresis revealed a main band with Mr = 46,000 both under reducing and non-reducing conditions. On gel filtration on Sephacryl S-300, however, the material was separated in two fractions, one eluting at the void volume, which contains active PAI-1, and one with Mr = 46,000 containing inactive material that could be reactivated with 12 M urea. SDS gel electrophoresis of the isolated high-Mr fraction revealed several bands including a main 46,000-Mr component, which reacted with anti-(PAI-1) antibodies on immunoblotting and neutralized tissue-type plasminogen activator (t-PA). The active high-Mr fraction and the reactivated low-Mr fraction of PAI-1 inhibited t-PA very rapidly with an apparent second-order rate constant of (1.5-4) x 10(7) M-1 s-1. The cDNA of endothelial cell PAI-1 was cloned and expressed in Chinese hamster ovary cells. The translation product, purified from conditioned medium of transfected cells, also revealed a high-Mr and a low-Mr fraction on gel filtration, which were indistinguishable from the natural proteins by physicochemical, immunochemical and functional analysis. On reduced SDS gel electrophoresis, the high-Mr fraction was separated into the Mr-46,000 low-Mr PAI-1 and two other components with Mr 65,000 and one barely entering the gel. When reactivated low-Mr PAI-1 was added to plasma, PAI activity and PAI-1 antigen eluted with an apparent Mr greater than or equal to 300,000 on gel filtration, indicating that active PAI-1 complexes with one or more binding proteins in plasma.

Cells, Cultured↗
PubMed1988

Biochemical and thrombolytic properties of a low molecular weight form (comprising Leu144 through Leu411) of recombinant single-chain urokinase-type plasminogen activator.

The cDNA encoding a low Mr derivative (residues 144-411) of human single-chain urokinase-type plasminogen activator was cloned, the recombinant low Mr single-chain urokinase-type plasminogen activator (rscu-PA-32k) was expressed in Chinese hamster ovary cells, and the translation product was purified to homogeneity from conditioned cell culture medium. rscu-PA-32k is very similar to intact recombinant single-chain urokinase-type plasminogen activator in terms of its very low activity (120 IU/mg) on a chromogenic substrate for urokinase (pyroglutamylglycylarginine p-nitroanilide), its plasminogen-dependent fibrinolytic activity on fibrin plates (specific activity = 170,000 IU/mg), its plasminogen activating potential, and the lack of specific binding to fibrin. In a rabbit jugular vein thrombosis model, comparable thrombolysis was obtained with rscu-PA-32k as compared to low molecular weight two-chain urokinase (50% lysis at 2.1 and 1.6 mg/kg infused over 4 h). Thrombolysis was associated with much less extensive systemic fibrinogen breakdown with rscu-PA-32k than with two-chain urokinase (residual fibrinogen at 50% lysis of 71 and 10%, respectively). It is concluded that the functional properties of rscu-PA-32k, expressed with a high efficiency, are similar to those of its previously characterized natural counterpart.

Animals↗
PubMed1988

Monitoring of hemostasis parameters during coronary thrombolysis with recombinant tissue-type plasminogen activator.

The monitoring of changes in the blood coagulation and fibrinolytic systems during thrombolytic therapy with recombinant tissue-type plasminogen activator (rt-PA) may be complicated by artifacts due to in vitro activation after blood collection and to interference of other agents (e.g., heparin) in the assays. In 106 patients with early acute myocardial infarction, infused with 150 mg of rt-PA (G11044) intravenously over 5 to 8 hours, blood samples were collected into liquid citrate supplemented with the plasmin inhibitor aprotinin (200 KIU/ml plasma) or on a lyophilized mixture of acidified citrate and the synthetic t-PA inhibitor D-Phe-Pro-Arg-CH2Cl (PPACK). A good correlation between precipitable (sulphite) and functional (clotting rate) fibrinogen levels was observed in plasma collected on citrate before therapy (r = 0.76) and in samples collected after 3 hours on either aprotinin (r = 0.87) or PPACK (r = 0.82). Precipitable fibrinogen levels were approximately 10% higher than functional level, in baseline samples collected on citrate alone and approximately 20% higher in 3 hour samples collected on either PPACK or aprotinin. Fibrinogen levels measured with both assays correlated well, but were somewhat higher in samples collected on PPACK than on aprotinin. rt-PA antigen levels assayed in plasma collected in either inhibitor correlated well (r = 0.90) but were 10-20% higher in PPACK containing samples. Addition of heparin up to 9 units/ml to plasma had no effect on the functional fibrinogen assay.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Chloromethyl Ketones↗
PubMed1988

Laboratory monitoring of hemostasis during thrombolytic therapy with recombinant human tissue-type plasminogen activator.

Recombinant tissue-type plasminogen activator (rt-PA) was administered intravenously to 93 patients with acute myocardial infarction and coronary thrombosis in doses of 30 to 150 mg over 1.5 to 6 hours. During this infusion plateau levels of rt-PA in plasma ranged between 0.4 and 2.2 micrograms/ml. Activation of the plasma fibrinolytic system and fibrinogen breakdown both in vivo and in vitro was observed with this therapy. In vitro fibrinogenolysis in plasma was more effectively prevented by collection of blood samples on aprotinin (200 kallikrein inhibitor units/ml blood), a conventional serine protease inhibitor, than on either of two monoclonal antibodies against t-PA (200 micrograms/ml plasma), or on D-phenylalanyl-prolyl-arginine-chloromethyl ketone (PPACK), a newly developed synthetic inhibitor of t-PA. Results of fibrinogen measurements during infusion of rt-PA were dependent on the method of assay. In a subgroup of 36 patients after completion of a thrombolytic infusion, fibrinogen decreased in vivo by 27% when measured as total coagulable protein and by 33% with a coagulation rate assay, but increased by 26% with an automated assay system. The extent of fibrinogenolysis was proportional to the plasma level of rt-PA but substantial intersubject variation was observed. Fibrinogenolysis in vivo was also associated with alpha 2-antiplasmin depletion and was more pronounced with a two-chain (G11021) than with a single-chain preparation (G11035) of rt-PA.

Fibrinogen↗
PubMed1988

Enzymatic properties of single-chain and two-chain forms of a Lys158----Glu158 mutant of urokinase-type plasminogen activator.

Recombinant single-chain urokinase-type plasminogen activator (rscu-PA), in which the plasmin-sensitive peptide bond Lys158-Ile159 is destroyed by site-specific mutagenesis of Lys158 to Glu (rscu-PA-Glu158), is quantitatively converted to two-chain urokinase-type plasminogen activator (rtcu-PA-Glu158) by treatment with endoproteinase Glu-C (Staphylococcus aureus V8 proteinase). The catalytic efficiency (k2/Km) of rscu-PA-Glu158 for the activation of plasminogen is 20 times lower (0.0001 microM-1 s-1) than that of rscu-PA (0.002 microM-1 s-1). In contrast, rtcu-PA-Glu158 has very similar properties to rtcu-PA obtained by digestion of rscu-PA with plasmin, including binding to benzamidine-Sepharose, catalytic efficiency for the activation of plasminogen (0.035 microM-1 s-1 versus 0.046 microM-1 s-1) and fibrinolytic activity in an in vitro plasma clot lysis system. It is concluded that the amino acid in position 158 is a main determinant of the functional properties of single-chain urokinase-type plasminogen activator but not of the two-chain form.

Catalysis↗
PubMed1988

New strategies in the development of thrombolytic agents.

Recombinant DNA technology has allowed large-scale production of the physiological, fibrin-specific, plasminogen activators tissue-type plasminogen activator (t-PA) and single-chain urokinase-type plasminogen activator (scu-PA). The results of clinical trials with these agents, mainly for the treatment of acute myocardial infarction, have revealed a limited fibrin specificity at the large therapeutic doses required for efficient thrombolysis. Mutants and variants of t-PA and scu-PA have given important information on structure-function relationships in these proteins and have resulted in rt-PA variants with significantly prolonged half-lives in vivo. Construction of chimaeric plasminogen activators containing various portions of t-PA and scu-PA has produced functionally active enzymes, however with a lower fibrin-affinity than wild-type t-PA. The promise of antibody targeting and the use of synergistic combinations of thrombolytic agents remains to be further investigated. We anticipate that eventually these research lines will yield artificial plasminogen activators with improved efficacy, risk/benefit and cost/benefit ratios.

Antibodies↗
PubMed1988

Detection of coronary artery reperfusion with creatine kinase-MB determinations during thrombolytic therapy: correlation with acute angiography.

Increases in plasma creatine kinase-MB (MB CK) were correlated with the onset of coronary artery reperfusion determined angiographically in 32 patients with acute myocardial infarction who were treated with recombinant human tissue-type plasminogen activator (rt-PA). Reperfusion occurred in 14 (70%) of 20 patients with left anterior descending coronary artery occlusion and in 8 (73%) of 11 patients with right coronary artery occlusion. One patient had persistent left circumflex coronary artery occlusion. Plasma MB CK levels (radioimmunometric assay) did not increase significantly in patients with persistent occlusion, but increased by a mean (+/- SEM) of 8 +/- 1 and 6 +/- 1 times over pretreatment levels at the end of the infusion in patients with a reperfused left anterior descending and right coronary artery, respectively. When a greater than or equal to 2.5-fold increase in MB CK levels at the end of the rt-PA infusion was taken as evidence of reperfusion of the left anterior descending coronary artery, 13 (93%) of 14 patients with reperfusion and 5 (83%) of 6 with persistent occlusion were correctly identified. When a greater than or equal to 2.2-fold increase in MB CK levels was used to identify right coronary artery reperfusion, seven (89%) of eight patients with persistent occlusion were correctly identified. The sensitivity and specificity of these indexes, derived from and applied to the same patient group, were 91 and 89%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Coronary Angiography↗
PubMed1988

Potential approaches for therapeutic intervention of thrombosis by fibrinolytic agents.

Fibrin-specific thrombolytic agents are being developed based on a better understanding of the molecular mechanisms that regulate in vivo fibrinolysis. These agents may be more effective than those available currently and may induce less systemic fibrinolysis. In this respect, t-PA has been extensively investigated. Another fibrinolytic substance with anticipated fibrin-selectivity is scu-PA. Although scu-PA has been much less extensively investigated than t-PA, sufficient knowledge is available to evaluate its potential as a fibrin-specific thrombolytic agent. Both t-PA and scu-PA are single-chain glycoproteins with a catalytic mechanism common to all serine proteases (active site composed of the charge relay system). Both molecules occur as single-chain forms but are converted easily to two-chain disulfide bonded molecules by digestion with plasmin. Unlike most other serine proteases for which the single-chain molecular form is a zymogen with little or no activity toward its natural substrate, both single-chain t-PA and scu-PA have inherent plasminogen-activating potential. Both t-PA and scu-PA induce fibrin-specific thrombolysis in a plasma environment and in animal models of thrombosis. However, the fibrin specificity of t-PA- and scu-PA-induced thrombolysis is based on different molecular mechanisms. The effectiveness and fibrin specificity of t-PA obtained by recombinant DNA technology recently were established by three randomized multicenter trials in patients with acute myocardial infarction. For scu-PA, clinical results are presently more limited.

Animals↗
PubMed1988

Pharmacokinetics and thrombolytic properties of a nonglycosylated mutant of human tissue-type plasminogen activator, lacking the finger and growth factor domains, in dogs with copper coil-induced coronary artery thrombosis.

The pharmacokinetics and thrombolytic properties of a variant of human tissue-type plasminogen activator (t-PA), obtained by deletion mutagenesis of the NH2-terminal fibronectin-like finger (F) and epidermal growth factor (E) domains, and substitution of the three known glycosylated Asn residues by Gln (t-PA-delta FE3X), were studied in dogs with a copper coil-induced thrombosis of the left anterior descending coronary artery. Bolus injections were given during 2 min to groups of three dogs. Injection of 0.15 mg/kg resulted in peak antigen levels in plasma of 1.58 +/- 0.72 micrograms/ml (mean +/- SEM) and caused reperfusion within 14 +/- 6 min. With 0.075 mg/kg, corresponding values of 0.81 +/- 0.20 micrograms/ml and 31 +/- 15 min were obtained. A bolus of 0.038 mg/kg yielded plasma peak levels of 0.43 +/- 0.20 micrograms/ml but did not cause coronary recanalization within 3 h. A bolus injection of natural t-PA (Mel-t-PA) at a dose of 0.1 mg/kg in four dogs resulted in plasma peak levels of 0.46 +/- 0.09 micrograms/ml and caused partial coronary artery reperfusion within 3 h in one of four dogs (after 31 min). None of these injections caused a significant decrease of the fibrinogen level. Pharmacokinetic parameters for t-PA-delta FE3X were alpha half-life (t1/2) 14-18 min, beta t1/2 72-125 min, and plasma clearance 21-36 ml/min. For Mel-t-PA, the corresponding values were 3 min, 8 min, and 520 ml/min. We conclude that the variant t-PA-delta FE3X has a markedly longer plasma t1/2 than does Mel-t-PA and, when administered as a bolus injection, a higher thrombolytic efficacy.

Animals↗
PubMed1988

Thrombolytic therapy.

Thrombolytic agents are plasminogen activators that convert plasminogen, the inactive proenzyme of the fibrinolytic system in blood, to the proteolytic enzyme plasmin. Plasmin in turn dissolves the fibrin of a blood clot, but may also degrade normal components of the hemostatic system and predispose to bleeding. The first-generation thrombolytic agents, streptokinase and urokinase, are only moderately efficacious, and their administration is associated with extensive systemic fibrinogen breakdown. In three major clinical trials in patients with acute myocardial infarction, early intravenous streptokinase significantly reduced in-hospital mortality. Tissue-type plasminogen activator (t-PA) is a more effective and fibrin-specific coronary arterial thrombolytic agent than streptokinase, but its impact on mortality remains to be established. Single-chain urokinase-type plasminogen activator (scu-PA, pro-urokinase) is more fibrin specific than urokinase but has only reached the stage of early clinical investigation. The acylated plasminogen streptokinase activator complex (Apsac) has a profile of thrombolytic efficacy and fibrin specificity that is probably very similar to that of streptokinase, but it can be administered as a single bolus injection. New trends toward further improved efficacy and fibrin specificity of thrombolytic therapy include the use of combinations of synergistic thrombolytic agents, mutants of t-PA or scu-PA, chimeric t-PA/scu-PA molecules, or antibody-targeted thrombolytic agents.

Clinical Trials as Topic↗