Abstract: A membrane filter of 0.025 to 0.05 .mu. in pore size is treated by passing the solution of a water-soluble high molecular substance such as albumin, dextran, polyvinylpyrrolidone, polysorbate 80, gelatin or the like through the membrane filter. Employing the filter thus treated, the solution of a physiologically active substance of human origin such as human growth hormone, kallikrein, trypsin inhibitor, epidermal growth factor, leucocyte interferon etc. is filtered at high recovery rate of the active substance avoiding the adsorption of the active substance onto the filter. By the filtration, harmful viruses such as Creutzfeldt-Jacob disease pathogen which may exist in the physiologically active substance can be removed.

Abstract: A method for the production of human tissue type plasminogen activator (tPa) using cells is disclosed. The method includes a supplementation of a p-aminomethyl benzoic acid derivative to a cell culture medium or a tPA producing medium and further an increase of osmotic pressure in the medium to 350 milliosmoles or more/liter. The invention provides a method for producing single-chain tPA in a high concentration and with a relatively small amount of double-chain tPA in the medium.

Abstract: Tissue plasminogen activator analogs exhibiting greater specificity for fibrin than native t-PA are disclosed. The analogs include the K1 domain of native t-PA replaced with another kringle domain mediating the binding of the analog to fibrin. The kringle contains six cysteine residues. The t-PA analogs may further include a variety of substitutions and modifications. Pharmaceutical compositions containing one or more of the t-PA analogs along with a physiologically acceptable carrier or diluent are also disclosed.

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: Chelating compounds of specified structure are useful for radiolabeling targeting molecules such as antibodies. Cleavable linkers connect the radionuclide metal chelates to the antibodies. The radiolabeled antibodies have improved biodistribution properties, including reduced localization within the intestines and kidneys.

Abstract: Disclosed is a means useful for selectively inhibiting human tissue plasminogen activator utilizing a thrombin inhibitor, illustrated by D-Phe-Pro-Arg-chloromethyl ketone (PPACK), thus providing a novel assay system for measuring human tissue plasminogen activator activity.

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: 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: The present invention provides a dry reagent for blood coagulation tests in which an at least partial course of the coagulation cascade takes place, comprising a carrier material which contains a chromophoric substrate of a protease of the blood coagulation system, at least one factor and/or co-factor of the blood coagulation system and a buffer.

Abstract: Disclosed herein are improved processes for preparing variant human t-PA proteins exhibiting improved pharmacokinetic properties relative to natural t-PA. One such illustrated variant, devoid of amino acids corresponding to amino acids 1 through 44 of natural t-PA, is shown to exhibit a plasma half-life of greater than about 15 times the plasma half-life to natural t-PA, as well as a clearance rate of less than about 1/10 the clearance rate of natural t-PA. Also disclosed are improved processes for treating vascular disease employing pharmaceutical compositions which incorporate therapeutically effective amounts of such t-PA variants with pharmaceutically acceptable diluents or excipients.

Abstract: An affinity gel for the isolation of serine proteases from a biological sample. The affinity gel is modified for specificity for serine proteases by coupling an organophosphorous compound to the gel. The reaction is carried out in two steps. The hydrophilic gel is reacted with a phosphoryl trifluoride and then the coupled gel is further reacted with an alcohol to form the organophosphorous compound.

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: Thrombolytic proteins are disclosed which have tissue plasminogen-type activity. The proteins are characterized by the presence of 0, 1, 2 or 3 N-linked polysaccharide substituents covalently bonded thereto and by a polypeptide sequence of the formula(a)A-R.sup.1 -B-R.sup.2 -C-R.sup.3 Dwherein A, B, C, and D are the following polypeptide sequences substantially as shown in FIG. 1:A is Gly-.sub.3 or Ser.sub.1 through Trp.sub.116, encompassing domain a;B is Ala.sub.120 through Gly.sub.183 ;C is Ala.sub.187 through Leu.sub.447 ;D is Val.sub.451 through Pro.sub.527 ; anda is a peptide domain comprising a sequence of 0-93 amino acids selected from the sequence Gly.sub.-3 or Ser.sub.1 through Thr.sub.91 ; R.sup.1, R.sup.2 and R.sup.3 are each a tripeptide sequence linking said A, B, C and D by peptide bonds, up to three of R.sup.1, R.sup.2 and R.sup.3 being a tripeptide sequence other than Asn-X-Thr or Asn-X-Ser, wherein X is any amino acid.

Abstract: A fibrinolytically active hybrid protein which comprises one chain of a 2-chain protease linked to a chain of a different 2-chain protease, or to the same chain of the same protease, at least one of the chains in the hybrid protein being derived from a fibrinolytically active protease, such that the hybrid protein has a catalytic site essential for fibrinolytic activity which is optionally blocked by a removable blocking group.

Abstract: A hybrid protein which comprises plasmin A-chain linked to urokinase B-chain, the catalytic site of which is blocked by a removable blocking group.

Abstract: A composition containing a tissue Plasminogen Activator (tPA) which comprises a partial hydrolyzate of gelatin cross-linked to a diisocyanate as an essential ingredient; or alternatively a partial hydrolyzate of gelatin cross-linked to a diisocyanate and one or more of a basic amino acid or salt thereof. The composition enhances the solubility of the tPA in water, thereby making the tPA further available in the treatment of circulatory diseases caused by thrombi.

Abstract: Fibrinolytic enzymes are produced and then isolated from non-cancerous established cell lines, designated as BEB or GPK. These fibrinolytic enzymes differ from those produced by malignant cell lines.

Abstract: Microplasmin and microplasminogen are plasmin and plasminogen derivatives produced by the action of plasmin non plasmin/plasminogen at high pH. The action of plasmin at this pH cleaves the heavy (A) chain of plasmin at the Arg.sub.529 - Lys.sub.503 or Lys.sub.530 -Leu.sub.531 bond and promotes disulfide bond rearrangement, producing microplasmin or microplasminogen consisting of a 30 or 31 residue C-terminal peptide derived from the A chain bound through new disulfide bonds to the intact B-chain of plasmin or plasminogen. The resulting products are significantly reduced in size and retain native activity.

Abstract: A fibrinolytically active hybrid protein which comprises one chain of a 2-chain protease linked to a chain of a different 2-chain protease, or to the same chain of the same protease, at least one of the chains in the hybrid protein being derived from a fibrinolytically active protease, such that the hybrid protein has a catalytic site essential for fibrinolytic activity which is optionally blocked by a removable blocking group.

Abstract: A derivative of a fibrinolytic enzyme in which the catalytic site on the enzyme which is responsible for fibrinolytic activity is blocked by a human protein attached thereto by way of a reversible linking group.

Abstract: The present invention provides a process for the determination of fibrin monomer in plasma, wherein a plasma sample containing fibrin monomer and freed from plasmin inhibitors is incubated in buffered solution with tissue plasminogen activator (EPA), plasminogen and a chromogenic plasmin substrate and the color formed is measured as a measure of the amount of fibrin monomer.The present invention also provides a reagent for the determination of fibrin monomer in plasma, wherein it contains plasminogen, tissue plasminogen activator (EPA), chromogenic plasmin substrate and buffer (pH 7.0 to 9.0).

Abstract: A fibrinolytically active protein conjugate comprising at least one optionally blocked fibrinolytic enzyme linked by way of a site other than the catalytic site responsible for fibrinolytic activity to at least one human protein.Processes from making the conjugates and pharmaceutical compositions containing them are also described.

Abstract: This invention relates to a method for determining fibrinolytic activation in human blood and to a method to distinguish between tissue (e.g. vascular) plasminogen activator and urokinase-like plasminogen activator contributions to fibrinolytic activation.Broadly, the method of the invention relates to detecting the level of plasmin-plasmin inhibitor complexes in blood plasma samples before and after clotting thereof; a greater increase in the complex level in the clotted sample as compared to non-clotted samples indicating the presence of circulating tissue plasminogen activator, with substantially identical levels of increase of complex in both the clotted and unclotted samples indicating the sole presence of urokinase-type plasminogen activator in the sample.

Abstract: A protease moiety having direct fibrinolytic activity which is useful in thrombolytic therapy is isolated from snake venom and shown to comprise a metalloproteinase having a molecular weight of from 25 to 27 kd and an isoelectric point of about 6.5 to 7.0. The moiety is separated from venom by a series of diverse fractionation steps including molecular sieve, ion-exchange and affinity chromatography.

Abstract: This invention relates to a method for producing a plasminogen preparation which has been pasteurized to produce a hepatitis safe injectable plasminogen. The method comprises: adding to an aqueous solution of plasminogen the protective agent of methyl lysine ester, or ethyl lysine ester or a hydrochloride salt thereof which has antifibrinolytic acitivity to thereby attach the agent to plasminogen and form a modified plasminogen; and subjecting the resulting modified agent to a heat treatment of at least 60.degree. C. for at least 10 hours. Further, the protective agent may be separated from the modified plasminogen by affinity chromatography.

Abstract: A derivative of a fibrinolytic enzyme in which the catalytic site on the enzyme which is responsible for fibrinolytic activity is blocked by a human protein attached thereto by way of a reversible linking group.

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: This invention provides a urokinase complex adsorbable by fibrin characterized in that it comprises heavy chain of high molecular weight urokinase as coupled with heavy chain of plasmin by one or more S-S bonds and a process for preparing the same.

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: Viruses, particularly hepatitis viruses, in blood clotting enzyme compositions are inactivated with little enzyme activity loss by heating the compositions in the dry state. The novel products which result are therapeutically and diagnostically useful.

Abstract: A process for quantitatively assaying factor Xa from a medium by reacting the medium with tripeptide derivatives having the formula ##STR1## wherein R.sup.5 is a chromogenic substituted amino group capable of being split off by enzymatic hydrolysis to form a colored or fluorescent product H--R.sup.5. The quantity of split product H--R.sup.5 released by the enzymatic action on the tripeptide derivative is determined photometrically, spectrophotometrically, fluorescence-spectrophotometrically, or electrochemically. The quantity of released split product H--R.sup.5 per time unit is proportional to the quantity of enzyme present in the starting material.

Abstract: A chromatographic procedure for the purification of a proteolytic procoagulant enzyme from extracts of human and animal tumors. The extracts are sequentially contacted with a first benzamide affinity chromatographic resin, an agarose filtration gel, a second benzamide affinity chromatographic resin and a phenyl-sepharose hydrophobic chromatographic resin. The resulting enzyme is capable of producing anti-procoagulant antibody which, which used in an immunoassay, is diagnostic for malignancy.

Abstract: What is disclosed is a process for rendering plasminogen virtually free of hepatitis virus by heating a plasminogen solution in the presence of a proteinase inhibitor with plasmin specificity, an amino acid, and a saccharide or sugar alcohol.

Abstract: A method for stabilizing plasminogen which comprises adding a physiologically acceptable inorganic salt to a plaminogen-containing aqueous solution to give a final concentration of the salt of 0.002 to 0.4 M.

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.

Abstract: What is disclosed is a glycoprotein having proteolytic, fibrinolytic, and thrombolytic properties and which can be used to prepare anti-sera useful as diagnostic agents, said glycoprotein being found in human blood serum and extracts of human placentas and being capable of isolation therefrom and which has the following properties:a protein proportion essentially consisting of 89.+-.4% of .alpha.-amino-acids,a carbohydrate proportion of 11.1.+-.2.2%, among it 5.3.+-.1.1% of hexoses, 2.8.+-.0.5 N of N-acetylated hexosamine, 2.9.+-.0.6% of N-acetylated neuraminic acid;a sedimentation coefficient S.sub.20x of 3.2.+-.0.3 S;a molecular weight of 32 000.+-.6 000;an iso-electric point at pH 4.3.+-.0.3;an extinction coefficient E.sub.1 cm.sup.1% (280 nm) of 13.8.+-.1.0;an electrophoretic mobility in the range between that of albumin and .alpha..sub.1 -globulins;a specific immunologic reaction with an antibody specifically directed against the protein, anda proteolytic activity, as well as a process for isolating it.