Abstract: Human elastase IV polypeptides and DNA (RNA) encoding such polypeptides and a procedure for producing such polypeptide by recombinant techniques and utilizing such polypeptide for therapeutic purposes, for example, restoration of elasticity of arterial walls, improvement of serum lipid abnormality and improvement of serum lipoprotein metabolism are disclosed. Also disclosed are antagonist/inhibitors against such polypeptides and their use in treating inflammation, arthritis, e.g. rheumatoid arthritis and osteoarthritis, septic shock, pancreatitis and limiting tissue damage in ulceration. Diagnostic assays for identifying mutations in nucleic acid sequence encoding a polypeptide of the present invention and for detecting altered levels of the polypeptide of the present invention are also disclosed.

Abstract: Mutants of the human PAI-1 protein are described which are inhibitors of neutrophil elastase or are inhibitors of vitronectin (Vn)-dependent cell migration. These mutants preferably comprise one or two amino acid substitutions in the reactive center loop of PAI-1, particularly at positions 331 and 346 of the mature protein. These mutants are notable in being resistant to inactivation by elastase, having high affinity for Vn, or both properties. These mutant proteins as pharmaceutical compositions are used to inhibit elastase in a subject, thereby treating a number of disorders associated with elastase activity, most notably emphysema, ARDS, inflammatory lung injury and cystic fibrosis. The mutants which interact with Vn are used to inhibit cell migration in a subject, thereby treating diseases or conditions associated with undesired cell migration and proliferation, particularly of smooth muscle cells.

Abstract: The present invention provides nucleic acid and corresponding amino acid sequences of a multifunctional protein that has been found to be useful in numerous medical and cosmetic contexts. A protein having "multifunctional activity," is defined herein as including at least one of a chymotrypsin, trypsin, collagenase, elastase or exo peptidase activity or asialo GM.sub.1 ceramide binding activity. These proteins are useful for multiple purposes, including treating viral infections such as herpes outbreaks, fungal, bacterial or parasitic infections, including the primary and secondary infections of leprosy, colitis, ulcers, hemorrhoids, corneal scarring, dental plaque, acne, cystic fibrosis, blood clots, wounds, immune disorders including autoimmune disease and cancer.

Abstract: A process is taught for the preparation and recovery of 7-aminodesacetoxycephalosporanic acid (7-ADCA) via enzymatic ring expansion activity on penicillin G, using a Penicillium chrysogenum transformant strain expressing expandase.

Abstract: A protein such as an enzyme or antibody is immobilized by crosslinking crystals of the protein with a multifunctional crosslinking agent. The crosslinked protein crystals may be lyophilized for storage. A preferred protein is an enzyme such as thermolysin, elastase, asparaginase, lysozyme, lipase or urease. Crosslinked enzyme crystals preferably retain at least 91% activity after incubation for three hours in the presence of a concentration of Pronase.TM. that causes the soluble uncrosslinked form of the enzyme to lose at least 94% of its initial activity under the same conditions. A preferred enzyme:Pronase.TM. ratio is 40:1. Enzyme crystals that are crosslinked may be microcrystals having a cross-section of 10.sup.-1 mm or less.

Abstract: Proteins such as enzymes and antibodies are immobilized by crosslinking crystals of the proteins such as microcrystals having a cross-section of 10.sup.-1 mm or less with a multifunctional crosslinking agent. The crosslinked protein crystals may be lyophilized for storage. Crystals of an enzyme such as thermolysin, elastase, asparaginase, lysozyme, lipase or urease may be crosslinked to provide crosslinked enzyme crystals that retain at least 91% activity after incubation for three hours in the presence of a concentration of Pronase.TM. that causes the soluble uncrosslinked form of the enzyme to lose at least 94% of its initial activity under the same conditions. A preferred Pronase.TM.:enzyme ratio is 1:40.

Abstract: A protein such as an enzyme or antibody is immobilized by crosslinking crystals of the protein with a multifunctional crosslinking agent. The crosslinked protein crystals may be lyophilized for storage. A preferred protein is an enzyme such as thermolysin, elastase, asparaginase, lysozyme, lipase or urease. Crosslinked enzyme crystals preferably retain at least 91% activity after incubation for three hours in the presence of a concentration of Pronase.TM. that causes the soluble uncrosslinked form of the enzyme to lose at least 94% of its initial activity under the same conditions. A preferred enzyme:Pronase.TM. ratio is 1:40. Enzyme crystals that are crosslinked may be microcrystals having a cross-section of 10.sup.-1 mm or less.

Abstract: Analogs of the serine protease inhibitors in which the amino acid sequence is varied slightly are disclosed, which analogs variously show improved properties including improved resistance to oxidative inactivation, improved ability to inhibit pancreatic elastase, improved ability to inhibit cathepsin G, and improved ability to inhibit trypsin.

Abstract: The invention is directed to a pharmaceutical preparation with a modulatory effect on malignant tumors, which contains a combination of protease proenzymes and amylases in a ratio between 1:100 and 100:1 in enzymatic activity units, and a protease inhibitor. The invention is also directed to use of such a preparation to modulate the effects of malignant tumors in humans and animals.

Abstract: Human elastase IV polypeptides and DNA (RNA) encoding such polypeptides and a procedure for producing such polypeptide by recombinant techniques and utilizing such polypeptide for therapeutic purposes, for example, restoration of elasticity of arterial walls, improvement of serum lipid abnormality and improvement of serum lipoprotein metabolism are disclosed. Also disclosed are antagonist/inhibitors against such polypeptides and their use in treating inflammation, arthritis, e.g. rheumatoid arthritis and osteoarthritis, septic shock, pancreatitis and limiting tissue damage in ulceration. Diagnostic assays for identifying mutations in nucleic acid sequence encoding a polypeptide of the present invention and for detecting altered levels of the polypeptide of the present invention are also disclosed.

Abstract: A protein such as an enzyme or antibody is immobilized by crosslinking crystals of the protein with a multifunctional crosslinking agent. The crosslinked protein crystals may be lyophilized for storage. A preferred protein is an enzyme such as thermolysin, elastase, asparaginase, lysozyme, lipase or urease. Crosslinked enzyme crystals preferably retain at least 91% activity after incubation for three hours in the presence of a concentration of Pronase.TM. that causes the soluble uncrosslinked form of the enzyme to lose at least 94% of its initial activity under the same conditions. A preferred enzyme:Pronase.TM. ratio is 1:40. Enzyme crystals that are crosslinked may be microcrystals having a cross-section of 10.sup.-1 mm or less.

Abstract: Detection or determination of a protease in a sample by incubating the sample with a substrate of the protease and observing proteolytic cleavage of said substrate. The substrate is a modified proenzyme containing a recognition site, e.g., an activation site, cleavable by said protease. Proteolytic cleavage of the modified proenzyme is detected by observing the resulting activity using a suitable substrate of the activated proenzyme. The protease may be e.g. an aspartic protease or a metalloprotease, and the modified proenzyme e.g. pro-urokinase having a mutant activation site which is cleavable by the protease to be determined.

Abstract: A protein such as an enzyme or antibody is immobilized by crosslinking crystals of the protein with a multifunctional crosslinking agent such as glutaraldehyde, and if desired lyophilizing the crosslinked crystals for storage. Crosslinking of the protein crystals provides stabilization for use under harsh conditions and for lyophilizing. The crystals crosslinked may be microcrystals having a cross-section of 10.sub.-1 mm or less. Crosslinked thermolysin, esterase, elastase, asparaginase and lysozyme crystals and crosslinked crystals of lipase from Geotrichum candidum and Candida cylindracea and of porcine origin can be used to convert a substrate to a product. Crosslinked thermolysin crystals are prepared that retain at least 96% of their initial activity after incubation for 4 days in the presence of a concentration of Pronase.TM. such as a thermolysin:Pronase.TM.

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: Methods are described for the identification and preparation of nucleic acid ligands to human neutrophil elastase. Included in the invention are specific RNA and DNA ligands to elastase identified by the SELEX method.

Abstract: The present invention provides for novel reagents whose fluorescence increases in the presence of particular proteases. The reagents comprise a characteristically folded peptide backbone each end of which is conjugated to a fluorophore. When the folded peptide is cleaved, as by digestion with a protease, the fluorophores provide a high intensity fluorescent signal at a visible wavelength. Because of their high fluorescence signal in the visible wavelengths, these protease indicators are particularly well suited for detection of protease activity in biological samples, in particular in frozen tissue sections. Thus this invention also provides for methods of detecting protease activity in situ in frozen sections.

Abstract: Human elastase IV polypeptides and DNA (RNA) encoding such polypeptides and a procedure for producing such polypeptide by recombinant techniques and utilizing such polypeptide for therapeutic purposes, for example, restoration of elasticity of arterial walls, improvement of serum lipid abnormality and improvement of serum lipoprotein metabolism are disclosed. Also disclosed are antagonist/inhibitors against such polypeptides and their use in treating inflammation, arthritis, e.g. rheumatoid arthritis and osteoarthritis, septic shock, pancreatitis and limiting tissue damage in ulceration.

Abstract: This invention particularly provides a novel polypeptide having high protease-inhibiting activity, preferably FXa-inhibiting activity, which comprises, at least as a part of the polypeptide, an amino acid sequence resulting from substitution of an amino acid for at least one amino acid in the following amino acid sequence (1), wherein the amino acid substitution is at least one substitution selected from the following substitution means (i) to (iii). It also provides a process for the production of the polypeptide, a novel DNA fragment encoding the polypeptide and a drug composition containing the same.Amino acid sequence (1) ##STR1## (i) Substitution of 15 position Gln counting from the N-terminus by an amino acid other than Gln.(ii) Substitution of 42 position Tyr counting from the N-terminus by an amino acid other than Tyr.(iii) Substitution of 7 position Arg counting from the N-terminus by an amino acid other than Arg.

Abstract: This invention provides a compound comprising:(1) a polypeptide moiety havinga) an identifying number of amino acids for SPAAT,b) an elastase binding activity; and(2) an extracellular matrix protein bound to the polypeptide moiety.Also provided is a method of inhibiting an elastase comprising contacting the elastase with a polypeptide moiety having:(1) an identifying number of amino acids for SPAAT;(2) a collagen binding activity; and(3) elastase binding activity.

Abstract: A new human elastase inhibitor is provided. The human monocyte elastase inhibitor is isolated, purified, characterized at the molecular level and cloned. The human monocyte elastase inhibitor is capable of forming a covalent complex with elastase or Proteinase-3 and is capable of inhibiting elastase.

Abstract: Aerosolization of therapeutic proteins is demonstrated using recombinant .alpha..sub.1 -antitrypsin for treatment of emphysema as paradigmatic.

Abstract: A protein such as an enzyme of antibody is immobilized by crosslinking crystals of the protein with a multifunctional crosslinking agent. The crosslinked protein crystals may be lyophilized for storage. A preferred protein is an enzyme such as thermolysin, elastase, asparaginase, lysozyme, lipase or urease. Crosslinked enzyme crystals preferably retain at least 91% activity after incubation for three hours in the presence of a concentration of Pronase.TM. that causes the soluble uncrosslinked form of the enzyme to lose at least 94% of its initial activity under the same conditions. A preferred enzyme:Pronase.TM. ratio is 1:40. Enzyme crystals that are crosslinked may be microcrystals having a cross-section of 10.sup.-1 mm or less.

Abstract: This invention particularly provides a novel polypeptide having high protease-inhibiting activity, preferably FXa-inhibiting activity, which comprises, an lease as a part of the polypeptide, an amino acid sequence resulting from substitution of an amino acid for at least one amino acid in the following amino acid sequence (1), wherein the amino acid substitution is at least one substitution selected from the following substitution means (i) to (iii). It also provides a process for the production of the polypeptide, a novel DNA fragment encoding the polypeptide and a drug composition containing the same.

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: Novel compounds which are chemically linked inhibitors of the proteases Elastase and Cathepsin G prevent connective tissue degradation associated with neutrophil induced inflammatory disease.

Abstract: This invention particularly provides a novel polypeptide having high protease-inhibiting activity, preferably FXa-inhibiting activity, which comprises, at least as a part of the polypeptide, an amino acid sequence resulting from substitution of an amino acid for at least one amino acid in the following amino acid sequence (1), wherein the amino acid substitution is at least one substitution selected from the following substitution means (i) to (iii). It also provides a process for the production of the polypeptide, a novel DNA fragment encoding the polypeptide and a drug composition containing the same.

Abstract: This invention provides a compound comprising:(1) a polypeptide moiety havinga) an identifying number of amino acids for SPAAT,b) an elastase binding activity; and(2) an extracellular matrix protein bound to the polypeptide moiety.Also provided is a method of inhibiting an elastase comprising contacting the elastase with a polypeptide moiety having:(1) an identifying number of amino acids for SPAAT;(2) a collagen binding activity; and(3) elastase binding activity.

Abstract: This invention relates to a novel protease-inhibitor,--which we called GELIN--and to pharmaceutical and cosmetic preparations thereof, containing this compound. GELIN is an inhibitor of human and porcine leucocyte elastase and chymotrypsin. GELIN has specific antibiotic properties. It also relates to the novel use of EGLIN, another chymotrypsin-inhibitor in cosmetic preparations.

Abstract: Substituted isocoumarins, their use in inhibiting serine proteases with trypsin-like, chymotrypsin-like and elastase-like specificity and their roles as anti-inflammatory agents.

Abstract: N-substituted azetidinones are a class of inhibitors of human leukocytes elastase which are known to be useful in the treatment of a wide variety of antiinflammatory and antidegenerative diseases. In inhibiting elastase, the therapeutic agents are shown to form a characteristic stable complex with the enzyme. In the assay disclosed herein, the inhibitor-enzyme complex is advantageously hydrolyzed and specific product(s) of the hydrolysis are measured. The assays are useful in a clinical setting, for determining appropriate dosage and assessing the effectiveness of treatment.

Abstract: One or more amino acid residues within the reactive site region of protease nexin-I are altered in order to create analogs or variants or protease nexin-I. These analogs have substantially different protease specificities as well as different effects on regulating the activity of proteolytic enzymes which enzymes have substantial effects on a number of different physiological functions. Formulations containing the protease nexin-I variants and methods for administering these formulations to obtain desirable therapeutic results are disclosed.

Abstract: The present invention provides a composition of matter useful as an antimicrobial agent. This composition of matter comprises an extract derived from human polymorphonuclear leukocytes, said extract including a polypeptide having an apparent molecular weight of about 13,000 daltons, a polypeptide having an apparent molecular weight of about 29,000 daltons and a polypeptide having an apparent molecular weight of about 54,000 daltons. The extract also has oxygen-independent, antimicrobial activity for bacteria and fungi at a pH from about 5.0 to about 8.0 and at calcium ion concentrations up to about 10 mM, bactericidal activity at sodium chloride concentrations up to about 0.3M, and fungicidal activity at sodium chloride concentrations up to about 0.15M. Methods for preparing this composition of matter are also provided. Further provided are purified polypeptides useful as antimicrobial agents, and methods for producing these polypeptides.

Abstract: The present invention provides a composition of matter useful as an antimicrobial agent. This composition of matter comprises at least two polypeptides selected from the group consisting of human polymorphonuclear leukocyte, polypeptides having on apparent molecular weights of about 3,500 daltons, about 13,000 daltons, about 18,000 daltons, about 29,000 daltons, and about 54,000 daltons. The composition of matter has respiratory burst-independent, antimicrobial activity for bacteria and fungi at a pH from about 5.0 to about 8.0 and at calcium ion concentrations up to about 10 mM, bactericidal activity at sodium chloride concentrations up to about 0.3M, and fungicidal activity at sodium chloride concentrations up to about 0.15M. Methods for preparing this composition of matter are also provided. Further provided are purified polypeptides useful as antimicrobial agents, and methods for producing these polypeptides.

Abstract: Human pancreatic elastase can now be obtained from a genetically engineered source.

Abstract: Human pancreatic elastase I can be obtained by genetic engineering.

Abstract: Certain novel heterocyclic compounds, aromatic thioesters, their preparation, and their use in inhibiting serine proteases with chymotrypsin-like and elastase-like specificity and in the treatment of diseases such as emphysema which involve tissue proteolysis.

Abstract: This invention relates to proteolytic enzymes with elastolytic properties from blood-sucking nematodes (hereinafter called HP enzymes). These enzymes are useful an anti-coagulants and as a means for dissolving fibrin clots. Because of their importance in the mechanism by which blood-sucking nematodes obtain nutrients from the host, HP enzymes would be an effective vaccine for prevention and treatment of these parasites. For the same reason, substances which inhibit HP enzymes would also be effective antithelminic agents.

Abstract: Certain novel aryl sulfonyl fluorides, their preparation, and their use in inhibiting serine proteases with chymotrypsin-like and elastase-like specificity.

Abstract: A process is disclosed for purifying elastase by precipitating elastase from an aqueous, elastase-containing solution, in which the electroconductivity of said aqueous solution is adjusted to 3 millimhos per centimeter or less and the pH of the solution is adjusted to a value of from 2 to 5. The electroconductivity of said aqueous solution is preferably adjusted to 3 millimhos per centimeter or less by means of ultrafiltration.

Abstract: An enzyme-containing detergent composition for a presterilization treatment of medical instruments and equipment, consisting of the following components, percent by weight:anionic surfactants: 4.0-6.0sodium phosphate: 30.0-35.0sodium silicate: 20.0-25.0sodium carbonate: 19.0-22.0soap comprising sodium salts of fatty acids: 2.0-4.0enzymatic preparation: 0.5-2.0sodium sulphate: the balance,the enzymatic preparation having the following composition, percent by weight:alkaline protease: 30-60neutral protease: 27-45elastase: 0.01-5.00collagenase: 0.001-4.00leucinaminopeptidase: 0.0001-0.011carboxypeptidase: 0.04-0.15fibrinolytic enzyme: 0.002-1.500lipase: 0.5-2.0amylase: the balance.

Abstract: An elastase-containing composition containing 0.5 to 50 parts by weight of sucrose fatty acid ester per part by weight of pure elastase, which composition permits elastase, useful as medicine for arterioscloerosis and hyperlipemia, to be absorbed in increased amount through intenstine.

Abstract: Protease is recovered by reacting a first amino acid whose amino group is protected with a protective group with a second amino acid whose carboxyl group is protected with a protective group, in an aqueous medium in the presence of a protease to result a peptide synthesis to deposit the addition compound of a dipeptide and said second C-terminal protected amino acid; dissolving said addition compound into the aqueous medium by adding a polar organic solvent which is miscible with water and separating an insoluble material from the resulting suspension by a solid-liquid separation to isolate the protease.

Abstract: Protease is recovered by reacting a first amino acid whose amino group is protected with a protective group with a second amino acid whose carboxyl group is protected with a protective group, in an aqueous medium in the presence of a protease to result a peptide synthesis to deposit the addition compound of a dipeptide and said second C-terminal protected amino acid; dissolving said addition compound by adding an organic solvent to a precipitate separated from the reaction mixture and isolating the protease from an organic solvent suspension or dissolving said addition compound by adding a water immiscible organic solvent and separating the organic solvent phase by a liquid-liquid separation.The addition compound can be dissolved in an organic solvent after separating the precipitate from the reaction mixture.The addition compound can be also dissolved by adding a water-immiscible organic solvent to the reaction mixture and the organic solvent phase can be separated from the aqueous phase.