Abstract: The present invention provides a vaccine to treat or prevent mastitis in a bovine species, comprising an immunologically effective amount of a plasminogen-activating streptokinase protein produced by Streptococcus uberis. The present invention further provides a method of treating or preventing mastitis in a bovine species comprising vaccinating a cow with an immunologically effective amount of a plasminogen-activating streptokinase protein produced by S. uberis. The present invention further provides an isolated plasminogen-activating streptokinase protein produced by S. uberis, or an immunogenic fragment thereof.

Abstract: A pharmaceutical preparation contains protein C and a thrombolytically active substance that does not activate protein C. This preparation prevents reocclusion usually occurring in the course of thrombolysis therapy.

Abstract: The invention relates to serine protease variants derived from precursor serine proteases via recombinant and/or chemical methods to form protease variants having improved peptide ligase activity. The invention also includes novel ligation substrates which in combination with the serine protease variants and a second ligation substrate are capable of forming a ligation product. The invention also relates to methods for forming such ligation products and the products formed thereby.

Abstract: rDSPA .alpha.1 is produced in commercial quantities and a purity adequate for clinical standards. The production methods utilize a series of chromatographic steps: cation exchange chromatography, followed by hydrophobic interaction chromatography, and ending with affinity chromatography.

Abstract: The invention relates to the use of thromboembolic proteins. These contain, as their only structural portion effecting thrombolytic activity, the protease domain of human tissue type plasminogen activator. These derivatives show reduced side effects, such as a reduction in bleeding while. demonstrating remarkable in vivo efficacy. The effect is surprising, given their in vitro properties.

Abstract: A multimolecular complex made up of a plasminogen activator conjugated to anti-ACE Mab 9B9 capable of delivering the plasminogen activator to the pulmonary endothelium is provided. Methods of using this complex to selectively deliver the plasminogen activator to the pulmonary endothelium to enhance fibrinolysis in the lungs of an animal are also provided. In addition, a method of prolonging the time a plasminogen activator is present in the circulation of an animal by conjugating the plasminogen activator to anti-ACE Mab 9B9 is also provided.

Abstract: This invention provides an improved method for enhancing the activity of thrombolytic agents, including t-PA, scu-PA, tcu-PA, streptokinase, acylated plasminogen-streptokinase activator complex (APSAC), mixtures of these, and other activators of plasminogen. The enhancement method comprises supplementation of plasma plasminogen levels with deglycosylated forms of glu- and lys-plasminogen. Deglycosylated plasminogen refers herein to glu- or lys-plasminogen 2 having a single oligosaccharide chain at Thr.sub.345, plasminogens having a single oligosaccharide chain at Asn.sub.288, and unglycosylated forms of plasminogen. The work described herein shows that a less glycosylated form of plasminogen (glu-plasminogen 2) has a higher affinity for fibrin clots than a more glycosylated plasminogen (glu-plasminogen 1).

Abstract: Nitrosylation of proteins and amino acid groups enables selective regulation of protein function, and also endows the proteins and amino acids with additional smooth muscle relaxant and platelet inhibitory capabilities. Thus, the invention relates to novel compounds achieved by nitrosylation of protein thiols. Such compounds include: S-nitroso-t-PA, S-nitroso-cathepsin; S-nitroso-lipoprotein; and S-nitroso-immunoglobulin. The invention also relates to therapeutic use of S-nitroso-protein compounds for regulating protein function, cellular metabolism and effecting vasodilation, platelet inhibition, relaxation of non-vascular smooth muscle, and increasing blood oxygen transport by hemoglobin and myoglobin. The compounds are also used to deliver nitric oxide in its most bioactive form in order to achieve the effects described above, or for in vitro nitrosylation of molecules present in the body.

Abstract: A method for preparing protease-polyethylene glycol adducts is presented wherein the immobilized reversible inhibitor, benzamidine, prevents reaction of activated polyethylene glycol with the active site of the protease. Improved activity against macromolecular substrates is obtained compared to when the benzamidine is in solution during the conjugation reaction.

Abstract: The invention involves thromboembolically effective compositions and therapeutic methods. Thrombolytically active proteins are combined with anticoagulants, as long as the anticoagulant is not heparin. The anticoagulant is administered in an intravenous bolus form, while the thrombolytically active protein may be administered via intravenous bolus, or intravenous infusion.

Abstract: Discussed are therapeutic approaches to the treatment of thrombolic conditions. The therapies use thrombolytically active proteins which inhibit reocclusion in the subject. The proteins are administered in two or more boli.

Abstract: Relatively inactive fusion proteins are activatable by enzymes of the clotting cascade to have fibrinolytic and/or clot formation inhibition activity. For example, a fusion protein comprising two hirudin or streptokinase molecules, linked by a cleavable linkage sequence, may be cleaved to yield anti-thrombotic hirudin or fibrinolytic streptokinase by thrombin or Factor Xa. Fibrinolytic or clot formation inhibition activity is therefore directed to the site of clot formation. Cleavable streptokinase/hirudin heterodimers are claimed.

Abstract: The method of preventing arterial thrombotic occlusion or thromboembolism by administering plasma-derived or recombinant produced protein C alone or in combination with a thrombolytic agent or combinations of thrombolytic agents.

Abstract: A process for the separation of streptokinase from contaminating proteins in a streptokinase-containing mixture, which comprises treating the mixture with a reducing agent to reduce disulphide bridges in the contaminating proteins to free thiol groups, contacting the mixture with a reagent R-X wherein R is a group capable of reacting with a free thiol group and X is a group R.sup.1 capable of reacting with a free thiol group or is a thiol-containing matrix, and thereafter separating the resulting chemically modified contaminating proteins from the mixture to provide streptokinase in a form substantially free of contaminating proteins.

Abstract: A pharmaceutical composition is prepared wherein a biologically active conjugated protein is selectively conjugated to at least one heparin fragment having a terminal 2,5-anhydro-D-mannose residue which has an aldehyde not involved in intramolecular hemiacetal formation. The resulting conjugate has a prolonged half-life as compared to native protein and is able to deliver heparin to the site of clots or to prevent reocclusion within the blood stream. Typical proteins include enzymes such as plasminogen activators, and in particular tissue plasminogen activators, and erythropoietin, hormones, antibodies and the like.

Abstract: The present invention relates to the field of biotechnology and genetic engineering and in particular a novel nucleotide sequence which codes for a streptokinase, as well as the recombinant DNA obtained therefrom which is used for the transformation of various host organisms.The present invention is based on the isolation of a new gene which codes for streptokinase from Streptococcus equisimilis of type C (strain ATCC-9542) and the cloning and expression thereof in prokaryotic (E. coli) and eukaryotic (Pichia pastoris) hosts, for which it includes the vehicles of expression which contain the genetic sequences of said gene, as well as the microorganisms transformed with these vectors capable of producing streptokinase.The protein obtained thereby can be used in clinical medicine as a therapeutic agent, in the treatment of disorders such as thromboembolic obstructions including coronary thrombosis.

Abstract: The present invention provides adducts of human t-PA derivatives, which comprise a t-PA derivative that lacks the Finger, Growth Factor and Kringle 1 domains, bound to an amphipathic molecule. The invention also provides methods for preparing the adducts and compositions for the treatment of thromboembolic disorders.

Abstract: The present invention relates to compositions comprising soluble complement receptor 1 (CR1) and a thrombolytic agent. In a specific embodiment, the thrombolytic agent is anisoylated human plasminogen-streptokinase activator complex (ASPAC). The invention further relates to methods for treating thrombotic conditions in humans and animals by administering a composition comprising soluble CR1 and a thrombolytic agent. In particular, the compositions and methods are useful both for reducing reperfusion injury and ameliorating the other effects of myocardial infarction.

Abstract: The present invention provides a method of treating thromboembolic disorders with the diglycosylated form of a tissue plasminogen activator derivative that lacks the Finger, EGF and Kringle I domains of the native tissue plasminogen activator molecule.

Abstract: This invention relates to a novel chemically synthesized gene including a base sequence coding for the primary amino acid sequence of natutal-type streptokinase, a corresponding plasmid recombinant, corresponding transformant and process for preparing streptokinase by the incubation of the transformant, the invention further relating to novel streptokinase derivative proteins having streptokinase activity and a modified primary amino acid sequence corresponding to the primary amino acid sequence of natural-type streptokinase which is deficient in the amino acid residues at the 373-position to the C-terminus, and wherein at least one of the amino acid residues may be deficient, replaced or inserted; the chemically synthesized gene including a base sequence coding for the derivative protein, plasmid containing the gene, transformant transformed by the plasmid, and process for preparing the streptokinase derivative protein by the incubation of the transformant.

Abstract: A screening method for the selection of mutagenized proteins that are normally secreted by cells is described. The method includes the development of a cloning vector for the expression of secretory proteins as fusion proteins on the cell surface of transfected mammalian cells. The secreted protein is displayed on the cell surface by fusion with the glycophospholipid membrane anchor of decay accelerating factor (DAF). Tissue-type plasminogen activator (t-PA), which is normally secreted, is used as a model protein. PCR mutagenesis is used to generate random mutations within the Kringle 1 (K1) domain of t-PA. Fluorescence activated cell sorting (FACS) is employed to screen for t-PA mutants possessing a loss of an epitope to a specific Mab, whose nonlinear binding domains overlap with the t-PA clearance receptor contact regions novel t-PA mutants designated N115S, N1425S, and K159R were discovered by this method.

Abstract: Light activated acyl-enzymes of the formula: ##STR1## are disclosed. In the compounds of Formula (III), ENZ is an enzyme, X is O or S, Y is --NR.sub.3 R.sub.4, --OR.sub.5, or --SR.sub.5, and Z is a nucleophile. m is 0 to 3 and n is 1 or 2. Y is substituted on the ring at either or both of the 4 and 6 position.R.sub.1 and R.sub.2 are each independently H, C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl.R.sub.3 and R.sub.4 are each independently H, C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl, except that R.sub.3 and R.sub.4 are not simultaneously both H. R.sub.5 is C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl.Methods of using the acyl-enzymes and intermediates for making the acyl-enzymes are disclosed.

Abstract: A cleavage-resistant plasminogen molecule is provided that is conveniently produced in recombinant cells by expression of a nucleic acid sequence encoding the plasminogen molecule. Preferably the plasminogen is a sequence variant with a modification in its two-chain cleavage site. The plasminogen molecule may be purified, acylated, complexed with acylated or non-acylated fibrinolytic enzymes, and formulated into pharmaceutical compositions for use in thrombolytic therapy.

Abstract: Compositions and methods of the synthesis of hybrid streptokinase with fibrin binding domains based on gene fusion technology. Recombinant DNA methods have been used to fuse gene segments for streptokinase and for fibrin-binding domains and express the fused gene in prokaryotic microorganisms, and the respective expressed protein is subsequently obtained by biotechnologic fermentation for the purpose of use in clinical medicine.

Abstract: A thrombolytic product comprising a fibrin-specific antibody substantially devoid of fibrinogen cross-reactivity coupled to a thrombolytic agent.

Abstract: Light activated acyl-enzymes of the formula: ##STR1## are disclosed. In the compounds of Formula (III), ENZ is an enzyme, X is O or S, Y is --NR.sub.3 R.sub.4, --OR.sub.5, or --SR.sub.5, and Z is a nucleophile. m is 0 to 3 and n is 1 or 2. Y is substituted on the ring at either or both of the 4 and 6 position.R.sub.1 and R.sub.2 are each independently H, C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl.R.sub.3 and R.sub.4 are each independently H, C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl, except that R.sub.3 and R.sub.4 are not simultaneously both H. R.sub.5 is C1 to C4 alkyl, C3 to C4 unconjugated alkenyl, or C3 to C4 unconjugated alkynyl.Methods of using the acyl-enzymes and intermediates for making the acyl-enzymes are disclosed.

Abstract: The method of preventing arterial thrombotic occulsion or thromboembolism by administering plasma-derived or recombinate produced protein C alone or in combination with a thrombolytic agent or combinations of thrombolytic agents.

Abstract: The present invention provides a method for the determination of a component of an immune reaction in a plasma sample using an immunoassay at a temperature of from 15.degree. to 40.degree. C., where one of the reactive components is in solid phase. The invention involves adding an amount of plasminogen activator sufficient to eliminate interference by fibrinogen with the component to be determined.

Abstract: A recombinant vector adapted for transformation of a suitable microorganism host to produce and secrete streptokinase. The recombinant vector comprises a plasmid into which a polydeoxyribonucleotide fragment from Streptococcus equisimilis H46A which codes for streptokinase synthesis and secretion has been inserted. The transformant microorganism including this recombinant plasmid vector produces streptokinase suitable for clinical fibrinolytic usage after purification.

Abstract: The thrombolytic effect of known plasminogen activators such as tissue plasminogen activator (t-PA), prourokinase and modifications of t-PA and prourokinase is enhanced dramatically and unexpectedly when combined with streptokinase. The body is treated with a combination streptokinase and one or more plasminogen activators of the t-PA or prourokinase type to achieve an enhanced effect which has advantage in thrombolytic therapy including the treatment of myocardial infarction.

Abstract: A process is described for the purification of plasminogen activator (PA), wherein a solution containing such as plasminogen activator is brought into contact with a carrier-bound polysulfate of a saccharide or sulfated sugar, the liquid is removed, and the PA bound by this material is eluted.

Abstract: Disclosed is a method of targeting reagents to the locus of a clot, and a family of substances that have affinity for fibrin. The method and substances exploit the discovery that the binding sites on protein A from Staphylococcus aureus have a significant affinity for fibrin. These fragments, analogs thereof, and oligomers of the fragments or analogs, may be attached to fibrinolytic enzymes, remotely detectable radiation emitting moieties, or healing agents to produce reagents which have affinity for the site of a wound or intravascular clot where fibrin has been deposited. Constructs comprising repeats of truncated analogs of the binding domain have low affinity for circulating immunoglobulin and high affinity for fibrin.

Abstract: A protein called PA binding protein has been isolated which binds specifically and reversibly to tissue plasminogen activator. The protein is characterized by a molecular mass of about 100,000 daltons, and electrophoretic mobility in agarose at pH 8.6 equal to that of plasma .beta.-globulins and an isoelectric point of 6.5 to 7.0. The protein is thermostable up to at least 56.degree. C. and is cleared from the circulation with a half life on the order of days.

Abstract: Disclosed herein is a method of enhancing the production of tissue plasminogen activator (t-PA) and single chain urokinase plasminogen activator (SCU-PA) by normal human lung diploid fibroblast cells in a serum-free medium. The method comprises the addition of heparin (or low molecular weight heparin) and an endothelial cell growth factor to the culture medium.

Abstract: A complex of a therapeutic agent is disclosed in which the therapeutic agent is complexed to an ammonium ion selected from the group consisting of ##STR1## where R.sup.1 and R.sup.2 are the same or different and are alkyl or hydroxy substituted alkyl containing 1 to 6 carbon atoms; andR, R' and R" are the same or different and are saturated or unsaturated aliphatics containing at least 10 carbon atoms, ##STR2## where R.sup.1, R.sup.2 and R" are as defined above, ##STR3## where R.sup.3 is independently alkyl or hydroxy substituted alkyl containing at least 10 carbon atoms; and R.sup.1, R.sup.2, R.sup.3 and R' having the meanings given above; and ##STR4## where R.sup.1, R.sup.2 and R.sup.3 have the meanings given above. The therapeutic agent is heparin, a biologically active peptide or protein or an antineoplastic drug. The therapeutic agent may also be covalently bonded to a triglyceride type backbone to form a compound.

Abstract: A binary complex between streptokinase and plasminogen is prepared in which the catalytic site essential for fibrinolytic activity is blocked by a group which is removable by hydrolysis such that the pseudo-first order rate constant for hydrolysis of the complex is in the range 10.sup.-6 sec.sup.-1 to 10.sup.-3 sec.sup.-1 in isotonic aqueous media at pH 7.4 at 37.degree. C. The complex is preferably a p-anisoyl streptokinase/plasminogen complex without internal peptide bond cleavage. The complex is useful in the treatment of venous thrombosis.

Abstract: A recombinant vector adapted for transformation of a suitable bacterial microorganism host to produce and secrete streptokinase. The recombinant vector comprises a plasmid into which a polydeoxyribonucleotide fragment from Streptococcus equisimilis H46A which codes for streptokinase synthesis and secretion has been inserted. The transformant bacterial microorganism including this recombinant plasmid vector produces streptokinase suitable for clinical fibrinolytic usage after purification.

Abstract: An aqueous solution of a tissue plasminogen activator dissolved therein at an increased concentration which comprises an aqueous medium and, dissolved therein, a high purity tissue plasminogen activator and at least one dissolution aid selected from the group consisting of lysine, ornithine and salts thereof, and a method for increasing a solubility of a high purity tissue plasminogen activator in an aqueous medium comprising adding said at least one dissolution aid to an aqueous solution of a high purity tissue plasminogen activator. The present aqueous solution contains a high purity tissue plasminogen activator dissolved therein at an increased concentration and the activity of the tissue plasminogen activator can be maintained during handling and storage of the aqueous solution.

Abstract: A method for stabilizing a tissue plasminogen activator which involves adding to an aqueous solution or powder containing a tissue plasminogen activator an effective amount of a purified gelatin is disclosed. A stable aqueous solution or powder which contains a tissue plasminogen activator and an effective amount of a purified gelatin is also disclosed. The method and composition eliminate the problem of adsorption of tissue plasminogen activator to various laboratory equipment, and also prevent the conversion of the single-chain tissue plasminogen activator into a double-chain tissue plasminogen activator.

Abstract: A derivative of streptokinase-human plasminogen activator complex, in which the active catalytic site essential for fibrinolytic activity is blocked by a 2- or 4-aminobenzoyl group, is useful in treating venous thrombosis.The blocking group is removable by hydrolysis such that the first order rate is in the range 0.7.times.10.sup.-5 sec.sup.-1 to 2.5.times.10.sup.-5 sec.sup.-1 in isotonic aqueous media at pH 7.4 at 37.degree. C.

Abstract: A process for providing enzyme activity to the surface of an article is described, comprising forming a film on the surface of the article, said film comprising an acid anhydride group-containing polymer and a polyol, wherein the acid anhydride groups are partially reacted with the polyol, and thereafter reacting the unreacted acid anhydride groups with an enzyme. Such an article provided with enzyme activity on the surface can be used for the production of food, medicines, etc., and as a medical material.

Abstract: A process for the preparation of a solid pharmaceutical or diagnostic composition in which the active substance and dextran are dissolved in water, the solution is placed in a mold and is thereafter lyophilized to form a solid abrasion-resistant tablet which can further be compressed to reduce its rate of disintegration in water. The invention is also directed to the pharmaceutical or diagnostic agent produced by this process.

Abstract: A pharmaceutical composition which comprises a pharmaceutically acceptable carrier together with an in vivo fibrinolytic enzyme as defined herein wherein the catalytic site essential for fibrinolytic activity is blocked by a group which is removable by hydrolysis at a rate such that the pseudo-first order rate constant for hydrolysis is in the range 10.sup.-6 sec.sup.-1 to 10.sup.-3 sec.sup.-1 in isotonic aqueous media at pH 7.4 at 37.degree. C.; is useful in the treatment of venous thrombosis.