Abstract: The present invention relates to physiologically-active derivatized natural and recombinant mammalian and human proteins and polypeptides. The invention provides chemical methods for derivatizing natural and recombinantly-derived proteins or polypeptides containing cysteine residues, either naturally or through site specific mutageneses. The pharmaceutical compositions containing the derivatized proteins and/or polypeptides are formulated to provide stable, long-acting compositions of such proteins and/or polypeptides, previously difficult to achieve.

Abstract: The present invention provides a method for screening candidate agents to identify compounds that modulate the hemostatic system. The method of the invention involves a screening medium comprised of stored whole blood, preferably diluted with buffer, to which unrefrigerated platelets have been added. Candidate agents that may inhibit or activate clot formation or clot lysis are added to the screening medium and suitable compounds are identified. The assay provided is physiologically relevant, rapid, inexpensive and allows for large scale screening of candidate agents.

Abstract: Polypeptides and fusion polypeptides that immunologically mimic the native Pl.sup.A1 and Pl.sup.A2 determinants of platelet protein GPIIIa are disclosed. Also disclosed are DNA segments and recombinant DNA molecules that encode those polypeptides and fusion polypeptides and methods for expressing and using those determinants. Antibodies that immunoreact with one determinant or the other, but not both are also disclosed, as are methods of preparing and using those antibodies.

Abstract: The present invention relates to a novel composition comprising coagulation Factor VIII and a non-ionic surfactant such as block co-polymers, e.g. polyoxamers or polyoxyethylene (20) fatty acid esters e.g. polysorbate 20 or polysorbate 80 as a stabilizer. The composition can also comprise sodium chloride, calcium chloride, L-histidine and/or sugars or sugar alcohols.

Abstract: A process for the preparation of factor IX from a biological source by chromatography involves prior treatment with ammonium sulfate as a protein precipitant at a concentration of from 1.5-2.3 mol/l.

Abstract: Disclosed is a method for treating liver injury caused by microorganism or toxic substance, which comprises administering to a patient suffering from liver injury a composition comprising a thrombomodulin, which has the ability to bind to thrombin and promote the activation of protein C by thrombin, as an active ingredient and at least one pharmaceutically acceptable carrier. The method of the present invention is very effective for ameliorating liver injury, such as fulminant hepatitis and hepatic veno-occlusive disease (VOD) which is likely to frequently occur after bone marrow transplantation.

Abstract: The present invention provides isolated DNA molecules comprising a DNA segment encoding novel human Kunitz-type inhibitors. Also provided are DNA constructs comprising a first DNA segment encoding a novel human Kunitz-type inhibitor wherein first DNA segment is operably linked to additional DNA segments required for the expression for the first DNA segment, as well as host cells containing such DNA constructs and methods for producing proteins from the host cells.

Abstract: Method for screening for the presence of a genetic defect associated with thrombosis and/or poor anticoagulant response to activated protein C (APC). The method is directed at detecting one or more mutations at one or more of the cleavage and/or binding sites for APC of Factor V and/or Factor Va or at Factor VIII and/or Factor VIIIa at either nucleic acid or protein level or both.

Abstract: A process for preparing thrombin which comprises treating a mixture comprising prothrombin, factor Xa, factor Va, and phospholipids with calcium ions, at a pH of 6.0-7.0 is provided. In particular the pH of 6.0-7.0 may be generated by the addition of the calcium ions or by buffering the preparation to a pH of 6.0-7.0. Thrombin preparations so produced may be subjected to further purification and are particularly stable even when substantially free of exogenous stabilizing agents such as proteins, sugars, polyol and mixtures thereof, and may be subject to freeze-drying and a virus inactivation by heat treatment.

Abstract: The invention relates to a pharmaceutical composition for the treatment of patients with blood coagulation diseases which are caused by coagulation factor deficiency and/or inhibitors of coagulation factors, whereby the composition has a FEIB-activity and is characterized in that it has Factor VIIa and at least one further active ingredient and the activity of at least 10 Factor VIIa units per unit FEIBA. Additionally, the invention comprises a method for the production of the pharmaceutical preparation and its use.

Abstract: A stable tissue adhesive is described which comprises fibrinogen and an activator or pro-activator of prothrombin, wherein its content of prothrombin present in blood is less than 5 units/g fibrinogen. This tissue adhesive can be present as a liquid or dry preparation and can optionally be applied to a biologically degradable water-soluble support.

Abstract: Disclosed are various compositions and methods for use in achieving specific blood coagulation. This is exemplified by the specific in vivo coagulation of tumor vasculature, causing tumor regression, through the site-specific delivery of a coagulant using a bispecific antibody.

Abstract: Method for screening for the presence of a genetic defect associated with thrombosis and/or poor anticoagulant response to activated protein C (APC). The method is directed at detecting one or more mutations at one or more of the cleavage and/or binding sites for APC of Factor V and/or Factor Va or at Factor VIII and/or Factor VIIIa at either nucleic acid or protein level or both.

Abstract: The invention provides high and low molecular weight fraction of von Willebrand Factor (vWF), which can be obtained by absorbing vWF to a heparin affinity support followed by eluting the vWF at differing salt concentrations The low molecular weight fraction is predominantly dimers and tetramers, and the high molecular weight fraction is predominantly larger multimers.

Abstract: This invention provides an imaging agent which comprises a polypeptide labeled with an imageable marker, such polypeptide having an amino acid sequence substantially present in the fibrin binding domain of naturally-occurring human fibronectin and being capable of binding to fibrin. The invention further provides a method wherein the imaging agent is used for imaging a fibrin-containing substance, i.e., a thrombus or atherosclerotic plaque. Further provided are plasmids for expression of polypeptides having an amino acid sequence substantially present in the fibrin binding domain of naturally-occurring human fibronectin and being capable of binding to fibrin, hosts containing these plasmids, methods of producing the polypeptides, methods of treatment using the polypeptides, and methods of recovering, refolding and reoxidizing the polypeptides.

Abstract: A clot lysis time determining device for determining the time necessary for fluid to lyse a clot. The device comprises: a body defining a chamber; a fluid receiving inlet port to the body in fluid communication with the chamber, the inlet port permitting fluid to flow into the chamber; a fluid discharging outlet port to the body in fluid communication with the chamber, the outlet port permitting fluid to flow out from the chamber; the inlet port, chamber, and outlet port defining a communicating fluid passageway; a body opening formed in the body along the fluid passageway, the body opening permitting a portion of fluid to flow against a clot supported by clot-supporting porous membrane; and a membrane cover detachably connectable to the body and configured for holding a clot-supporting porous membrane over the body opening.

Abstract: The present invention provides methods, reagents, and test kits for evaluating the effect of a test composition on APC response by employing factor Va.sup.R506Q Methods and compositions for treating patients suffering from reduced APC response are also provided.

Abstract: The catalytic active site of Factor VII is modified to produce a compound which effectively interrupts the blood coagulation cascade. The modification renders Factor VIIa substantially unable to activate plasma Factors X or IX. Pharmaceutical compositions of the modified Factor VII are used to treat a variety of coagulation-related disorders.

Abstract: Recombinant rabbit tissue factor is cloned and produced in a bacterial host. This protein, which is relatively insoluble and has several disulfide bonds, requires special modifications in order to express and be purified at commercial levels. By expressing the tissue factor as a fusion protein with a bacterial enzyme, thioredoxin, solubility is increased. Use of a thioredoxin reductase deficient host aids in proper tertiary structure for biological activity.

Abstract: The present invention is directed to immunochemical detection procedures, e.g., using both Western blotting and direct immunoassays, for fragments of Factor Va, which can thus be used; (a) in a predictive manner to evaluate the existence and/or extent of a thrombotic complication; (b) to monitor the efficacy of prophylaxis for a thrombotic condition; and (c) as a means to evaluate potential risk of hemorrhage during thrombolytic therapy.

Abstract: The DNA encoding the cell surface receptor for thrombin has been cloned and sequenced. The availability of this DNA permits the recombinant production of thrombin receptor which can be produced at cell surfaces and is useful in assay systems both for the detection of thrombin and for the evaluation of candidate thrombin agonists and antagonists. Further, the elucidation of the structure of the thrombin receptor permits the design of agonist and antagonist compounds which are useful diagnostically and therapeutically. The availability of the thrombin receptor also permits production of antibodies specifically immunoreactive with the receptor per se or with specific regions thereof which are also useful diagnostically or therapeutically.

Abstract: The invention relates to a method for isolation of highly pure von Willebrand Factor in which recombinant von Willebrand Factor (rvWF) is chromatographically purified by anion exchange chromatography on an anion exchanger of the quaternary amino type in a buffer solution comprising buffer substances and optionally salt. The buffer solutions are preferably free of stabilizers, amino acids and other additives. According to this method, highly pure recombinant rvWF can be obtained, which is free from blood plasma proteins, especially free from Factor VIII, and is physiologically active. Further, the invention relates to a pharmaceutical preparation that contains rvWF, which comprises mulitimers with a high structural integrity.

Abstract: The invention provides compounds which specifically inhibit factor Xa activity. The compounds consist of the structure X.sub.1 -YIR-X.sub.2, wherein X.sub.1 is H, acyl, alkyl, acylalkyl, arylalkyl or one or more amino acids, and X.sub.2 is a modified C-terminal group, one or more carboxy-protecting groups or one or more amino acids or other substituent, and Y, I and R are tyrosine, isoleucine and arginine, respectively, or peptidomimetic or organic structures that possess the same functional activity as Y, I and R, respectively. In addition, the present invention provides a compound having the structure A1-A2-(A3).sub.m --B, where m is 0 or 1. A compound of the invention can be linear or cyclic and can be about 2 and 43 residues in length. A compound of the invention is characterized, in part, in that it exhibits a specific inhibition of factor Xa activity with a K.sub.i of .ltoreq.100 .mu.M, preferably .ltoreq.

Abstract: Narrative complement pathway proteins modified such that the protein is capable of forming a stable C3 convertase. Preferably the modified protein is a modified human C3 protein. DNA sequences encoding such proteins are also provided, together with DNA constructs. Conjugates comprising such proteins and a specific binding moiety, for example an antibody, are also described, as are uses of such proteins and/or conjugates in therapy.

Abstract: Modified Protein C molecules have been made which substitute the gamma carboxylglutamic acid (Gla) region of another Vitamin K dependent protein, most preferably prothrombin, for the native region of the Protein C. A modified protein C molecules has been made which substitutes the gamma carboxyglutamic acid (Gla) region with the corresponding region of prothrombin. The modified or chimeric protein C has advantages over the wild-type protein C since it is less sensitive to inhibition by some natural antibody inhibitors of protein C (which would otherwise decrease the ability of the protein C to act as an anticoagulant) and which do not need the same cofactors or same amounts of cofactors, and can therefore be effective in patients with lowered levels of the cofactors such as protein S or the lipids present in elevated levels in platelets such as phosphatidyl ethanolamine (PE). The anticoagulant activity of the chimera was tested in normal and factor V Leiden plasma.

Abstract: A method and device for activating and applying a solution of fibrinogen to a desired site. The method includes contacting the solution of fibrinogen with immobilized thrombin resulting in an activated solution of polymerizable fibrin, and delivering the activated solution to the desired site. The device includes a housing having a compartment for a solution of fibrinogen, immobilized thrombin, a structure for bringing the solution of fibrinogen in contact with the immobilized thrombin under conditions which permit the activation of the fibrinogen resulting in polymerizable fibrin, and a structure for delivery of the activated solution to the desired site.

Abstract: Modified Protein C molecules have been made which substitute the gamma carboxylglutamic acid (Gla) region of another Vitamin K dependent protein, most preferably prothrombin, for the native region of the Protein C. The modified or chimeric protein C has advantages over the wild-type protein C since it is less sensitive to inhibition by natural inhibitors of protein C (which would otherwise decrease the ability of the protein C to act as an anticoagulant) and which does not need the same cofactors or same amounts of cofactors, and can therefore be effective in patients with lowered levels of the cofactors such as protein S or the lipids present in activated platelets such as phosphatidyl ethanolamine (PE).

Abstract: Analogs of blood factors which are transiently inactive are useful in treatment of diseases characterized by thrombosis. In addition, modified forms of activated blood factors that generate the active blood factor in serum but have extended half-lives are useful in treating hemophilic conditions. These modified forms of the blood factor may be acylated forms which are slowly deacylated in vivo.

Abstract: The present invention is directed to a soluble thrombomodulin-containing composition comprising at least one molecular species of soluble thrombomodulin, and at least one member selected from the group consisting of maltose, lactose, sucrose, arginine and a salt thereof, and a nonionic surface-active agent; and a production method therefor; as well as a stabilizing agent; a stabilization method; an anti-adsorption agent; and an anti-adsorption method for the soluble thrombomodulin. The present invention provides a soluble thrombomodulin-containing lyophilized preparation which is useful as a prophylactic or therapeutic agent for diseases associated with abnormalities in blood coagulation, and which is stable for a prolonged period of time and would not become adsorbed onto the container; and a production method therefor; as well as a stabilization agent; a stabilization method; an anti-adsorption agent; and an anti-adsorption method for the soluble thrombomodulin.

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 concerns a method for inhibiting the growth and/or causing regression of tumors by administering a therapeutically effective dose of a procoagulant and a cytokine, preferably TNF-.beta., TNF-.alpha. and/or IL-1. In a specific aspect, the invention concerns a method for tumor treatment by the administration of a therapeutically effective amount of a thrombomodulin inhibitor and a cytokine. The invention also concerns thrombomodulin inhibitors and pharmaceutical compositions used in the course of these treatments.

Abstract: Hybrid proteins having cross-linking and tissue-binding activities, DNA molecules encoding such proteins and methods for producing the hybrid proteins from recombinant host cells are disclosed. The hybrid proteins disclosed herein are useful in tissue sealant and wound healing formulations.

Abstract: A method and a device for separating a component, such as fibrin monomer from blood, by centrifugation, involve feeding of blood admixed an anticoagulant to a first annular chamber in a device, where the annular chamber is defined by a cylindrical outer wall and a cylindrical inner wall, both walls extending coaxially about a common axis, as well as by a top wall and a bottom wall. The top wall or the bottom wall is formed by a piston body displaceable within the first chamber. This method involves furthermore a centrifugation of the device about the said common axis to substantially separate blood into a cell fraction and a plasma fraction followed by the resulting plasma fraction being transferred while influenced by the piston body to a second chamber defined by an outer cylindrical wall. The outer cylindrical wall extends coaxially with the said common axis, whereby a fraction with fibrin I is caused to be separated in the second chamber while a suitable enzyme is being added.

Abstract: The invention relates to fibrin sealants. More specifically, the invention relates to the preparation of a fibrin polymer wherein the polymer is substantially free of the enzyme.

Abstract: The invention lies in the field of genetic engineering and, in particular, is concerned with the use of 7B2 as chaperone in vivo or in vitro. The invention accordingly concerns a method for producing a desired protein in vivo with the aid of recombinant cells capable of expressing 7B2 and of expressing and secreting said desired protein. Another aspect is accordingly an in vitro method for the deaggregation or prevention of aggregation of protein by treating the protein with 7B2.

Abstract: Reagents that specifically bind a carbohydrate target wherein sialic acid is linked at the nonreducing terminus of a glycoside to a galactose or galactosamine residue through an .alpha.2-6 linkage are able to inhibit the conversion of human Factor X to human Factor Xa. These reagents (SA/Gal/GalNAc binding reagents) as well as other strategies for inhibiting the conversion of Factor X to Factor Xa are useful in treating thrombosis, inflammation and other conditions associated with excess thrombin activity.

Abstract: A process for viral inactivation of a solution containing a biologically active protein, wherein the process comprises the steps of 1) contacting the solution with an immobilized ligand under conditions which allow protein to bind to the ligand, 2) subjecting the bound protein to a viral inactivation method under conditions which would result in substantial denaturation of the protein if it were not bound to the ligand, and 3) recovering the protein by washing the immobilized protein under conditions which favor the release of the protein into the solution under conditions in which the recovered protein retains its biological activity.

Abstract: A process for producing a highly purified preparation of an inhibited form of an activated blood factor entails providing a partially purified preparation containing the blood factor of interest, treating the partially purified preparation to convert the blood factor to an inhibited activated form in a single step, and then purifying the resulting inhibited activated blood factor.

Abstract: Enhanced production of cryoprecipitate is produced by dehydrating an individual unit of plasma prior to a low temperature step used to produce cryoprecipitate. This dehydration is accomplished either by placing a water absorbing material within a blood bag so that plasma occupying the bag will become dehydrated or by placing the water absorbing material within a cartridge so that plasma becomes dehydrated upon flowing through the cartridge. The preferred water-absorbing material is a cross-linked chromatographic gel having pores too small to admit clotting proteins, but large enough to admit water molecules. Suitable gels are made from carbohydrates or polyacrylamide. Carbohydrate gels such as Sephadex.RTM., produced by Pharmacia-Upjohn, are particularly preferred in the present invention.

Abstract: The invention relates to a pharmaceutical preparation comprising a plasma protein wherein said preparation is free of infectious agents as well as essentially free of denaturation products and is obtainable by a method that encompasses the following steps:a) addition of a polyether and a chaotropic agent to a solution comprising the plasma protein, optional lyophilization of the solution;b) inactivation of infectious agents in the presence of the polyether by a physio-chemical or chemical treatment, andc) removal of the polyether and the chaotropic agent.

Abstract: The disclosed Factor V Ratio (FVR) screening blood assay (read as "factor five ratio") and kits for the conduct thereof, identify individuals that possess a specific genetic defect, known as the Factor V Leiden defect, or other genetic or acquired Factor V defect, that makes those individuals susceptible to venous thromboembolism. In this test the Factor V activity of a blood plasma sample exposed to activated Protein C (APC) is compared to the Factor V activity of a similar sample in the absence of APC, after both samples had been treated with an activating agent. The ratio between the Factor V activity level without APC and the Factor V activity level with APC, identifies individuals at risk of a thrombotic disorder due to a Factor V defect and differentiates between individuals with a heterozygous defect and individuals with a homozygous defect.

Abstract: The present invention describes methods for inhibiting angiogenesis in tissues using vitronectin .alpha..sub.v .beta..sub.3 antagonists, and particularly for inhibiting angiogenesis in inflamed tissues and in tumor tissues and metastases using therapeutic compositions containing .alpha..sub.v .beta..sub.3 antagonists.

Abstract: The invention concerns a method for inducing a selective collapse of the vasculature of a solid tumor by administering to a patient a therapeutically effective dose of a combination of a compound preventing the formation of a functional thrombin-thrombomodulin complex and a cytokine selected from the group of TNF-.beta. (LT), TNF-.alpha., IL-1, and IFN-.gamma.. The invention further concerns the composition used in this method.

Abstract: Novel kits for the preparation of a fibrin sealant are disclosed. In one kit a first component is fibrin monomer which can be polymerized to form a fibrin sealant when combined with the second component which is distilled water or an alkaline buffer.

Abstract: The subject invention relates to methods and compositions using nondynamic fibrin monomer which is selectively converted to a fibrin polymer which serves as the fibrin sealant.

Abstract: An assay for activated factor VII (factor VIIa) has been developed using truncated tissue factor (tTF), a soluble mutant form of tissue factor (TF) that retains the cofactor function of TF toward factor VIIa. Unlike full-length TF, however, tTF appears not to support the conversion of factor VII to VIIa. As a result, the tTF assay for factor VIIa is free from interference from factor VII in the plasma and is therefore specific for factor VIIa. The assay is much simpler than existing assays, because it is a single-stage clotting assay performed almost identically to a prothrombin time (PT) assay. It is also considerably more sensitive than current assays for factor VIIa in plasma. Since the tTF assay is calibrated against a factor VIIa standard, it yields an absolute concentration of factor VIIa in ng/ml.

Abstract: A tissue factor protein mutant capable of neutralizing the ability of endogenous tissue factor to induce coagulation is provided. A representative tissue factor mutant designated K165A, K166A TF is useful in a method for inhibiting thrombin-induced platelet aggregation in a mammal, either separately or in combination with a thrombolytic agent, an anticoagulant, or a GPII.sub.b III.sub.a inhibitor.

Abstract: The invention relates to fibrin sealants. More specifically, the invention relates to the use of a fibrin sealant wherein a composition comprising fibrin monomer or a composition comprising noncrosslinked fibrin is utilized as a component of the fibrin sealant.

Abstract: The invention provides compositions having a Factor VIIa active site inhibitor domain and a tissue factor domain for the inhibition of FVIIa. The invention also provides pharmaceutical compositions comprising the novel compositions as well as their use in diagnostic, therapeutic, and prophylactic methods.

Abstract: The present invention relates to a method for detecting disturbances of the protein C/protein S system.