Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.73 (urokinase-type plasminogen activator)
 10,685 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

K1K2Pu, a recombinant t-PA/u-PA chimera with increased thrombolytic potency in animal models of venous and arterial thrombosis, which consists of amino acids 1 to 3 and 87 to 274 of human tissue-type plasminogen activator (t-PA) and amino acids 138 to 411 of human single chain urokinase-type plasminogen activator (scu-PA), was produced and conditioned for use in patients. Chinese hamster ovary cells were transfected with an expression plasmid containing the K1K2Pu cDNA, high producer cell lines were selected and scaled up in 800 cm2 roller bottles, and 350 ml conditioned cell culture medium was harvested 3 to 7 times at 2 to 5 day intervals. Batches of 21 +/- 4 liter (mean +/- SD, n = 28) containing 1.8 +/- 0.6 mg/l of K1K2Pu related antigen were purified by chromatography on Copper chelate-Sepharose and immunoadsorption on an insolubilized murine monoclonal antibody (MA-1C8). Yields were 8.6 +/- 3.4 mg K1K2Pu per batch with a specific activity of 83,000 +/- 44,000 IU/mg. The final material, obtained at a concentration of approximately 0.7 mg/ml, was dialyzed against 0.3 M NaCl, 0.02 M Tris-HCl buffer, pH 7.5, containing 0.01% Tween 80 and 10 KIU/ml aprotinin. It was homogeneous on SDS-PAGE, contained 6.5 +/- 6.9 percent two chain material and the contamination with murine monoclonal antibody was less than 0.1 percent. After filtration of pools of 3 to 5 selected batches on 0.22 microns Millipore filters the material was sterile and virus free by routine screening; it was obtained at a concentration of approximately 0.5 mg/ml with a specific activity of 110,000 +/- 16,000 IU/mg (mean +/- SD, n = 3) and an endotoxin content of 0.5 to 7 units/mg. Bolus injection at a dose of 1 mg/kg in mice did not produce weight loss within 8 days. Thus, this material appears to be suitable for the investigation on a pilot scale of the pharmacokinetic and thrombolytic properties of K1K2Pu in patients with thromboembolic disease.
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PMID:K1K2Pu, a recombinant t-PA/u-PA chimera with increased thrombolytic potency, consisting of amino acids 1 to 3 and 87 to 274 of human tissue-type plasminogen activator (t-PA) and amino acids 138 to 411 of human single chain urokinase-type plasminogen activator (scu-PA). Purification in centigram quantities and conditioning for use in man. 163 5

Expression of plasminogen activator inhibitor 2 (PAI-2) under the control of the protease B gene promoter in a mutant strain of Saccharomyces cerevisiae, DS569, resulted in its accumulation intracellularly at up to 20% of the soluble cell protein. Provision of an N-terminal signal sequence resulted in the secretion of a hyperglycosylated molecule. The intracellularly produced PAI-2 was purified by copper-chelate and anion-exchange chromatography to greater than 95% pure and was fully active. The recombinant PAI-2 formed SDS-stable complexes with urokinase and tissue-type plasminogen activator and inhibited the proteases with similar reaction kinetics to placental PAI-2 (second-order rate constant for uPA, 2.4 x 10(6) M-1 s-1, and for two-chain tPA, 0.7 x 10(5) M-1 s-1). As is the case for placental PAI-2, the N-terminus of the yeast-derived recombinant PAI-2 was blocked. The high productivity and consequent ease of purification mean that S. cerevisiae provides an excellent source of recombinant PAI-2 for investigation of its therapeutic potential in the treatment of neoplastic and inflammatory diseases.
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PMID:Purification and characterisation of plasminogen activator inhibitor 2 produced in Saccharomyces cerevisiae. 190 Oct 39

The thrombolytic efficacy of recombinant unglycosylated full length single chain urokinase-type plasminogen activator (rscu-PA, saruplase), applied either as single intravenous bolus or as a continuous infusion over 60 minutes, was studied in 5 randomized blinded groups of 5 dogs with combined copper coil induced coronary artery thrombosis and 125I-fibrin labeled femoral vein clots. Infusion of 1 mg/kg recu-PA (group I) induced coronary recanalization in 4 of 5 dogs and 98 +/- 1% (mean +/- SEM) venous clot lysis. Bolus injection of 1 mg/kg recu-PA (group II) caused reflow in 3 of 5 dogs and 88 +/- 5 percent venous clot lysis. Infusion of 0.5 mg/kg rescu-PA (group III) achieved reflow in 3 of 5 dogs and 52 +/- 6% venous clot lysis. Bolus injection of 0.5 mg/kg rscu-PA (group IV) induced reflow in 4 of 5 dogs and 48 +/- 12% venous clot lysis. Placebo infusion (group V) was associated with late recanalization in 1 of 5 dogs and 18 +/- 8% venous clot lysis. Coronary artery reocclusion after reflow was not observed in groups I and II, but occurred in 2 of 3 animals in group III and in 3 of 4 animals in group IV (P = .02). The time to reflow in responsive animals was 22 +/- 5 minutes with infusion of 0.5 or 1 mg/kg rscu-PA and 14 +/- 1 minute with bolus injection of 0.5 or 1 mg/kg (P = .14). Depletion of fibrinogen and alpha 2-antiplasmin to less than 25% of baseline levels was observed in the 5 dogs given 1 mg/kg rscu-PA by bolus and in 3 of the 5 dogs given 1 mg/kg rscu-PA via infusion, but in none of the dogs that received 0.5 mg/kg rscu-PA (P less than .001). Plasma clearance rates were 170 +/- 44 and 230 +/- 30 mL/minute after bolus injection and 190 +/- 47 and 310 +/- 56 mL/minute during infusion of rscu-PA for the 1 mg/kg and 0.5 mg/kg doses respectively. Thus, intravenous bolus injection of rscu-PA (saruplase) appears to be equipotent to an infusion over 60 minutes for both coronary and venous thrombolysis. This animal model of combined arterial and venous thrombolysis may be useful for the evaluation of new thrombolytic strategies.
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PMID:Comparison of intravenous bolus injection or continuous infusion of recombinant single chain urokinase-type plasminogen activator (saruplase) for thrombolysis. A canine model of combined coronary arterial and femoral venous thrombosis. 211 30

The dose-response of an intravenous i.v. infusion for 30 min of recombinant human single-chain urokinase-type plasminogen activator (rscu-PA) was investigated in dogs with 1-h-old clots in the left anterior descending coronary artery. The clots were induced with a copper coil, and thrombolysis was monitored by repeated coronary angiography. Intravenous infusion of rscu-PA at a rate of 2 micrograms/kg/min did not induce lysis within 30 min (n = 4). Infusion at a rate of 4 micrograms/kg/min in 7 dogs produced complete lysis in 2 (within 25 and 27 min), partial lysis in 2 (within 18 and 25 min), and no lysis in 3. Infusion at 8 micrograms/kg/min in four dogs caused complete lysis in three dogs within 18 +/- 3 min (mean +/- SD) and partial lysis in the fourth animal. Infusion at 20 micrograms/kg/min in four dogs induced complete lysis within 14 +/- 3 min. A linear correlation was observed between the infusion rate and the plateau level of rscu-PA in blood. At the highest infusion rate (20 micrograms/kg/min), the concentration of rscu-PA in blood was 2.5 +/- 0.45 microgram/ml, but this was not associated with systemic fibrinolytic activation because the alpha 2-antiplasmin and fibrinogen levels remained essentially unchanged. It is concluded that i.v. infusion of recombinant single-chain urokinase-type plasminogen activator (rscu-PA) at a sufficiently high rate (greater than or equal to 8 micrograms/kg/min) produces coronary thrombolysis without systemic fibrinolysis in dogs.
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PMID:Thrombolysis with recombinant human single-chain urokinase-type plasminogen activator (rscu-PA): dose-response in dogs with coronary artery thrombosis. 243 1

An instrument consisting of a copper vapor laser coupled to an optical fiber/chemical injector catheter for the treatment of occluded arteries has been constructed and tested. The combined application of three steps: the pre-irradiation injection of a light absorbing dye, HPD; brief copper laser irradiation (at 578 nm); and a urokinase infusion after the irradiation, produced the striking effect of liquefaction and resolution of thrombus. The histological examination of the arteries after the treatment showed no apparent damage to the arterial wall.
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PMID:Copper vapor laser and optical fiber catheter system for liquefaction and removal of thrombus in occluded arteries. 256 71

The data on tumour vascularization regulating substances, their structure and mechanism of action are reviewed. The systems for testing angiogenic reactions are analyzed. The role of mast cells, lymphocytes, macrophages and leucocytes in formation of new microvessels is under discussion. The results of the search for antitumor drugs inhibiting angiogenesis are promising. The data are analyzed on the role of prostaglandins, Cu2+ and urokinase in the processes of tumour vascularization. The possibility of the complex and multifactor regulation of microvessels is suggested.
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PMID:[Regulation of angiogenesis and tumor growth]. 257 91

S-S cross-linking enzyme, skin sulfhydryl oxidase (SSO), catalyzes the formation of disulfide bonds from sulfhydryl groups in skin. The activity of SSO was detected in differing amounts in each of the four layers--stratum corneum, stratum granulosum, stratum spinosum with basal cell layer, and dermis--of cow snout skin, with the highest specific activity being recorded in the stratum granulosum. SSO was stimulated to 130-150% of its initial activity by treatment with 1 mg/ml trypsin, chymotrypsin, or urokinase, but was not affected by plasmin or cathepsin D. These findings suggest that SSO may be activated by some kinds of serine proteases during the keratinocyte autolysis process in the stratum granulosum. SSO showed the highest activity with the addition of 5 microM of Cu2+. The atomic absorptive analysis of purified SSO showed 0.5 atoms of Cu in one molecule of SSO. From these findings, it was determined that Cu2+ was essential for the activity of SSO. The molar ratio of the disappearance of DTT, consumption of O2, and production of H2O2 during the enzyme reaction was 1:1.05:0.89. From these findings, the reactions catalyzed by SSO is suggested to be represented by the following equation: (table; see text).
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PMID:[Localization in skin, activation and reaction mechanisms of skin sulfhydryl oxidase]. 258 80

The effect of pretreatment with heparin on coronary thrombolysis by plasminogen pro-activator was studied in 20 dogs with a 1-h old clot in the left anterior descending coronary artery. The clot was induced by placement of a copper coil and thrombolysis was confirmed angiographically. Intravenous administration of plasminogen pro-activator at a rate of 5 micrograms/kg/min (group 1; n = 5) and 20 micrograms/kg/min (group 2; n = 5) after pretreatment with heparin (300 IU/kg) induced thrombolysis in 31 +/- 4 min and 14 +/- 2 min, respectively. When plasminogen pro-activator was administrated intravenously at a rate of 20 micrograms/kg/min without heparin pretreatment (group 3; n = 5), the infusion interval for successful thrombolysis was prolonged to 45 +/- 6 min, which was significantly longer than that in groups 1 and 2 (p less than 0.001). In these three treatment groups, thrombolysis was not associated with severe alteration in plasma hemostatic factors (fibrinogen and alpha 2-antiplasmin). An evaluation of plasma urokinase activities using a chromogenic substrate S-2444 did not show that heparin increased the plasma urokinase activities. By increasing the dose of plasminogen pro-activities. By increasing the dose of plasminogen pro-activator to 80 micrograms/kg/min, successful reperfusion was rapidly obtained in 25 +/- 5 min without heparin pretreatment (group 4; n = 5); this was significantly faster than the results seen in group 3 (p less than 0.001). An analysis of urokinase activities showed that plasminogen pro-activator was fully converted to urokinase, which induced complete depletion of fibrinogen and alpha 2-antiplasmin.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Enhancement of coronary thrombolysis with plasminogen pro-activator by pretreatment with heparin. 312 1

Experimental studies in dogs with coronary thrombi induced by copper wire confirmed the optimal method of intracoronary thrombolysis, and showed that a high-dose, brief intravenous infusion of urokinase can lead to recanalization. The thrombolytic effects of intracoronary thrombolysin at a rate of 50 IU/kg over 10 minutes are similar to the effects of intracoronary urokinase at a rate 500 IU/kg over 20 minutes. Overall reperfusion rates of 83-86% have been achieved. These results indicate that the thrombolytic effect of thrombolysin is 20 times stronger than that of urokinase. The effect of a brief intravenous infusion of urokinase was less than that of intracoronary urokinase. The reperfusion rate in the same experimental model was 40%. Later, a clinical trial of intracoronary urokinase was performed in 47 patients with acute myocardial infarction who were admitted within 12 hours of the onset of symptoms. In 25 of 33 (75.8%) patients with complete occlusion, selective or ostial infusion of urokinase 500 IU/kg over 20 minutes was successful. When given intravenously, recanalization was achieved in 11 of 15 (73%) patients with complete occlusion who were admitted within 6 hours. Both reperfusion rates were similar except for dosage and the duration of infusion.
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PMID:Indications for and limitations of coronary thrombolysis. 316 42

The thrombolytic efficacy of defibrase (DF) and urokinase (UK) was evaluated and compared in a canine model in which left circumflex coronary artery (LCX) thrombus formation was initiated by electrical stimulation (150 UA, DC) of the arterial intimal surface via an implanted copper wire electrode. Forty-eight mongrel dogs were equally divided into 8 groups: six groups for both intravenous (iv) and intracoronary (ic) infusion of DF, UK and normal saline 30 min after complete obstruction by a LCX thrombus, the remaining two groups for iv infusion DF and UK 360 min after occlusion. Results showed that in the control groups no LCX recanalization after infusion occurred, but all the treated groups recanalized when drug infusion started 30 min after occlusion. Thrombus wet weight and infarct size were much higher in the control group. No significant differences were found between treated groups except that the recanalizing speed was fastest by ic UK with the highest occurrence rate of reperfusion arrhythmia. For 6-hour-old thrombi, DF was more effective than UK for recanalization. Thrombus wet weight and infarct size were lower in the recanalized dogs. Both agents reduced plasma plasminogen and fibrinogen significantly but serious incision bleeding was only found in the UK group. In order to study the mechanism underlying UK-induced hemorrhage, we observed the influences of UK and DF on blood coagulation factors in another 12 dogs. Results showed that DF only reduced factor 1 while UK additionally reduced coagulation factors 2, 7, 8 and 10, indicating that the non-selective depletive effect on coagulation factors may be one of the mechanisms of UK-induced hemorrhage. We conclude that both DF and UK can lyse fresh coronary thrombi, DF was more effective than UK on lysing older thrombi, considering its convenient use and less frequency of side effects, DF is therefore considered a more promising agent clinically.
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PMID:Comparison of the thrombolytic efficacy of defibrase and urokinase on canine coronary artery thrombosis and the mechanism of urokinase-induced hemorrhage. 349 37


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