Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues
Patent
1997-11-24
2000-07-18
Wax, Robert A.
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
C07K 100
Patent
active
060909161
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to specific proteins as well as recombinant versions of these proteins which are serine protease inhibitors, including potent anticoagulants in human plasma. These proteins include certain proteins extracted from nematodes. In another aspect, the present invention relates to compositions comprising these proteins, which are useful as potent and specific inhibitors of blood coagulation enzymes in vitro and in vivo, and methods for their use as in vitro diagnostic agents, or as in vivo therapeutic agents, to prevent the clotting of blood. In a further aspect, the invention relates to nucleic acid sequences, including mRNA and DNA, encoding the proteins and their use in vectors to transfect or transform host cells and as probes to isolate certain related genes in other species and organisms.
BACKGROUND AND INTRODUCTION TO THE INVENTION
Normal hemostasis is the result of a delicate balance between the processes of clot formation (blood coagulation) and clot dissolution (fibrinolysis). The complex interactions between blood cells, specific plasma proteins and the vascular surface, maintain the fluidity of blood unless injury occurs. Damage to the endothelial barrier lining the vascular wall exposes underlying tissue to these blood components. This in turn triggers a series of biochemical reactions altering the hemostatic balance in favor of blood coagulation which can either result in the desired formation of a hemostatic plug stemming the loss of blood or the undesirable formation of an occlusive intravascular thrombus resulting in reduced or complete lack of blood flow to the affected organ.
The blood coagulation response is the culmination of a series of amplified reactions in which several specific zymogens of serine proteases in plasma are activated by limited proteolysis. This series of reactions results in the formation of an insoluble matrix composed of fibrin and cellular components which is required for the stabilization of the primary hemostatic plug or thrombus. The initiation and propagation of the proteolytic activation reactions occurs through a series of amplified pathways which are localized to membranous surfaces at the site of vascular injury (Mann, K. G., Nesheim, M. E., Church, W. R., Haley, P. and Krishnaswamy, S. (1990) Blood 76: 1-16. and Lawson, J. H., Kalafatis, M., Stram, S., and Mann, K. G. (1994) J. Biol. Chem. 26: 23357-23366).
Initiation of the blood coagulation response to vascular injury follows the formation of a catalytic complex composed of serine protease factor VIIa and the non-enzymatic co-factor, tissue factor (TF) (Rappaport, S. I. and Rao, L. V. M. (1992) Arteriosclerosis and Thrombosis 12: 1112-1121). This response appears to be exclusively regulated by the exposure of subendothelial TF to trace circulating levels of factor VIIa and its zymogen factor VII, following a focal breakdown in vascular integrity. Autoactivation results in an increase in the number of factor VIIa/TF complexes which are responsible for the formation of the serine protease factor Xa. It is believed that in addition to the factor VIIa/TF complex, the small amount of factor Xa which is formed primes the coagulation response through the proteolytic modification of factor IX to factor IX.sub.alpha which in turn is converted to the active serine protease factor IXa.sub.beta by the factor VIIa/TF complex (Mann, K. G., Krishnaswamy, S. and Lawson, J. H. (1992) Sem. Hematology 22: 213-226.). It is factor IXa.sub.beta in complex with activated factor VIIIa, which appears to be responsible for the production of significant quantities of factor Xa which subsequently catalyzes the penultimate step in the blood coagulation cascade; the formation of the serine protease thrombin.
Factor Xa catalyzes the formation of thrombin following the assembly of the prothrombinase complex which is composed of factor Xa, the non-enzymatic co-factor Va and the substrate prothrombin (factor II) assembled in most cases, on the surface of activated platelets which are ad
REFERENCES:
patent: 4458066 (1984-07-01), Caruthers et al.
patent: 4683293 (1987-07-01), Craig
patent: 4745051 (1988-05-01), Smith et al.
patent: 4777242 (1988-10-01), Nelles
patent: 4808537 (1989-02-01), Stroman et al.
patent: 4812405 (1989-03-01), Lair et al.
patent: 4818700 (1989-04-01), Cregg et al.
patent: 4837148 (1989-06-01), Cregg
patent: 4855231 (1989-08-01), Stroman et al.
patent: 4857467 (1989-08-01), Sreekrishna et al.
patent: 4879231 (1989-11-01), Stroman et al.
patent: 4882279 (1989-11-01), Cregg
patent: 4885242 (1989-12-01), Cregg
patent: 4895800 (1990-01-01), Tschopp et al.
patent: 5002876 (1991-03-01), Sreekrishma et al.
patent: 5004688 (1991-04-01), Craig et al.
patent: 5023236 (1991-06-01), Edgington et al.
patent: 5032516 (1991-07-01), Cregg
patent: 5106833 (1992-04-01), Broze et al.
patent: 5122465 (1992-06-01), Cregg et al.
patent: 5135868 (1992-08-01), Cregg
patent: 5166329 (1992-11-01), Cregg
patent: 5189019 (1993-02-01), Palladino et al.
patent: 5204261 (1993-04-01), Prevatt et al.
patent: 5239058 (1993-08-01), Vlasuk et al.
patent: 5239059 (1993-08-01), Zasloff et al.
patent: 5268273 (1993-12-01), Buckholz
patent: 5330901 (1994-07-01), Prevatt et al.
patent: 5427937 (1995-06-01), Cappello et al.
patent: 5525477 (1996-06-01), Hassouna
patent: 5605671 (1997-02-01), Lyle et al.
Aoki, Y., et al., "Effects of Recombinant Human Soluble Thrombomodulin (rhs-TM) on a Rat Model of Disseminated Intravascular Coagulation with Decreased Levels of Plasma Antithrombin" Thrombosis and Hemostasis 71(4):452-455 (1994).
Babin et al., "The Isoinhibitors of Chymotrypsin/Elastase from Ascaris lumbricoides: The Primary Structure" Arch. of Biochem. and Biophy. 232(1):143-161 (1984).
Beaucage et al., "Deoxynucleoside Phosphoramidites--A New Class of Key Intermediates for Deoxypolynucleotide Synthesis" Tetrahedron Letters, 22(20):1859-1862 (1981).
Bernard, et al., "The Serine Protease Inhibitor Family from Ascaris Suum: Chemical Determination of the Five Disulfide Bridges" Arch. Biochem. Biophys., 303(2):367-376 (1993).
Bock, P.E. et al. "Isolation of Human Coagulation a-Factor X.sub.a by Soybean Trypsin Inhibitor-Sepharose Chromatography and Its Active-Site Titration with Fluorescein Mono-p-guanidinobenzoate" Archives of Biochem. Biophys. 273(2):375-388 (1989).
Bolivar et al., "Construction and Characterization of New Cloning Vehicles" Gene, 2:95-113 (1977).
Broach, J. et al., "Transformation In Yeast: Development of a Hybrid Cloning Vector and Isolation of the CAN1 Gene" Gene, 8:121-133 (1978).
Brown, E. et al., "Chemical Synthesis and Cloning of a Tyrosine tRNA Gene" Methods in Enzymology, 68, 109-151 (1979).
Bullock et al., "XL1-Blue: A High Efficiency Plasmid Transforming recA Escherichia coli Strain with Beta-Galactosidase Selection" Biotechniques 5(4):376-379 (1987).
Cairns et al., "Antithrombotic Agents in Coronary Artery Disease" Chest 102:456S-481S (1992).
Crameri et al., "Display of Biologically Active Proteins on the Surface of Filamentous Phages: a cDNA Cloning System for Selection of Functional Gene Products Linked to the Genetic Information Responsible for their Production" Gene, 137:69-75 (1993).
Cappello et al., "Ancylostoma Factor Xa Inhibitor: Partial Purification and Its Identification as a Major Hookworm-Derived Anticoagulant In Vitro" J. Infect. Diseases, 167:1474-1477 (1993).
Cappello et al., "Ancylostoma caninum anticoagulant peptide: A hookworm-derived Inhibitor of Human Coagulation Factor Xa," Proc. Natl. Acad. Sci. U.S.A. 92:6152-6156 (1995).
Carroll et al., "The Anticoagulant Effects of the Hookworm, Ancylostoma Ceylanicum: Observations on Human and Dog Blood In Vitro and Infected Dogs In Vivo" Thromb. Haemostas. (Stuttgart), 51(2):222-227 (1984).
Carson, "Computerized Analysis of Enzyme Cascade Reactions Using Continuous Rate Data Obtained with an Elisa Reader" Comput. Prog. Biomed 19:151-157 (1985).
Clements et al., "Secretion of Human Epidermal Growth Factor from Saccharomyces Cerevisiae Using Synthetic Leader Sequences" Gene
Bergum Peter W.
Gansemans Yannick Georges Jozef
Jespers Laurent Stephane
LaRoche Yves Rene
Lauwereys Marc Josef
Corvas International Inc.
Wax Robert A.
LandOfFree
Nematode-extracted serine protease inhibitors and anticoagulant does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Nematode-extracted serine protease inhibitors and anticoagulant , we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Nematode-extracted serine protease inhibitors and anticoagulant will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2037974