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(12) Patent Application Publication (10) Pub. No.: US 2003/0180934A1 Ni Et Al

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(12) Patent Application Publication (10) Pub. No.: US 2003/0180934A1 Ni Et Al US 2003O180934A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2003/0180934A1 Ni et al. (43) Pub. Date: Sep. 25, 2003 (54) PLASMINOGEN-LIKE POLYNUCLEOTIDES, (60) Provisional application No. 60/158,044, filed on Oct. POLYPEPTIDES, AND ANTIBODIES 7, 1999. (75) Inventors: Jian Ni, Germantown, MD (US); Paul Publication Classification E. Young, Gaithersburg, MD (US); Steven M. Ruben, Olney, MD (US) 51) Int.nt. Cl.C.7 ............................ C12N 9/64; CO7H 21/04 C12P 21/02; C12N 5/06 Correspondence Address: (52) U.S. Cl. ..................... 435/226; 435/69.1; 435/320.1; HUMAN GENOME SCIENCES INC 435/325; 536/23.2 941O KEY WEST AVENUE ROCKVILLE, MD 20850 (73) Assignee: Human Genome Sciences, Inc., Rock (57) ABSTRACT ville, MD (US) (21) Appl. No.: 10/162,742 The present invention relates to novel human plasminogen like polypeptides and isolated nucleic acids containing the (22) Filed: Jun. 6, 2002 coding regions of the genes encoding Such polypeptides. Related U.S. Application Data Also provided are vectors, host cells, antibodies, and recom binant methods for producing human plasminogen-like (63) Continuation of application No. 09/832,197, filed on polypeptides. The invention further relates to diagnostic and Apr. 11, 2001, now abandoned, which is a continua therapeutic methods useful for diagnosing and treating dis tion-in-part of application No. PCT/US00/27253, orders related to these novel human plasminogen-like filed on Oct. 4, 2000. polypeptides. US 2003/0180934 A1 Sep. 25, 2003 PLASMINOGEN-LIKE POLYNUCLEOTIDES, the inactive Zymogen plasminogen, which is hydrolyzed to POLYPEPTIDES, AND ANTIBODIES yield the active two-chain Serine protease plasmin. Plasmin is a member of the chymotrypsin Superfamily of proteins CROSS-REFERENCE TO RELATED which use Serine as the nucleophile in peptide bond cleav APPLICATONS age. Plasminogen is activated to form plasmin by the 0001. This application is a continuation of and claims hydrolysis of the arg560-val561 peptide bond in plasmino priority under 35 U.S.C. S 120 to U.S. application Ser. No. gen. This hydrolysis is performed by the naturally occurring 09/832,197, filed Apr. 11, 2001, which is a continuation-in "plasminogen activators' urokinase plasminogen activator part of, and claims benefit under 35 U.S.C. S120 to Inter (u-PA) and the tissue-type plasminogen activator (t-PA), as national Application No. PCT/US00/27253, filed Oct. 4, well as by Streptokinase and Staphylokinase proteins. 2000, which claims benefit under 35 U.S.C. S 119(e) of U.S. 0006. In addition to fibrinolysis, plasmin has been impli Provisional Application No. 60/158,044, filed Oct. 7, 1999, cated in Several other physiological processes Such as extra all of which are hereby incorporated by reference in their cellular-matrix degradation, tissue remodeling, cell migra entireties. tion, neuronal cell migration, inflammation, angiogenesis, and wound healing (Bugge, T. H. et al., Cell, 87:709-19 FIELD OF THE INVENTION (1996)). Likewise, it is thought that plasmin has a broad 0002 The present invention relates to novel plasmino Substrate Specificity, as it is known to degrade Several gen-like proteins. More specifically, isolated nucleic acid common extracellular-matrix glycoproteins in vitro (Bugge, molecules are provided encoding novel plasminogen-like T. H. et al., (1996)). polypeptides. Novel plasminogen-like polypeptides and 0007. The activation of plasminogen to plasmin is tightly antibodies that bind to these polypeptides are provided. Also regulated by the activities of Serine protease inhibitors, Such provided are vectors, host cells, and recombinant and Syn as plasminogen activator inhibitor (PAI) type-1, which thetic methods for producing human plasminogen-like poly inhibit the activities of the plasminogen activators u-PA and nucleotides and/or polypeptides. The invention further t-PA (Chapman, H. A. (1997)). PAI-1, for example, is a relates to diagnostic and therapeutic methods useful for glycoprotein of approximately 50 kd that has been impli diagnosing, treating, preventing and/or prognosing disorders cated as the cause of reduced fibrinolytic capacity of plasma related to these novel plasminogen-like polypeptides. The from Survivors of myocardial infarctions (Hamsten, A. et al., invention further relates to Screening methods for identify New Eng. J. Med., 313:1557-1563 (1985); Pannekoek, H. et ing agonists and antagonists of polynucleotides and al, EMBO.J., 5:2539-2544 (1986); Ginsberg, D. et al., J. Clin. polypeptides of the invention. The present invention further Invest., 78:1673-1680 (1980)). relates to methods and/or compositions for inhibiting the production and function of the polypeptides of the present 0008 Thus there exists a clear need for identifying and invention. exploiting novel members of the plasminogen-like chymot rypsin family of Serine proteases. Although Structurally BACKGROUND OF THE INVENTION related, Such proteins may possess diverse and multifaceted functions in a variety of cell and tissue types. The purified 0.003 Hemostasis is maintained through the opposing plasminogen-like polypeptides of the invention are research forces of fibrinolysis and blood coagulation. The formation tools useful for the identification, characterization and puri of blood clots is part of the body's natural response to injury fication of additional plasminogen activating molecules. and trauma. Blood clot formation is mediated through the Furthermore, the identification of new Serine proteases per coagulation pathway, which produces the prothrombotic mits the development of a range of derivatives, agonists and enzyme thrombin. Thrombin converts circulating fibrinogen antagonists at the nucleic acid and protein levels which in to fibrin, which is an insoluble polymer that forms the turn have applications in the treatment and diagnosis of a framework of a blood clot. The resulting blood clot serves to range of conditions Such as cancer, inflammation, angiogen arrest the flow of blood from the site of injury or trauma, esis, cell migration, ECM degradation, tissue remodeling, Such as a cut or other tissue wound (H. A. Chapman, Curr: wound healing, neurological disorders and blood clotting Biology, 9:714-24 (1997)). disorders, amongst other conditions. 0004 Although beneficial in promoting the arrest of blood flow from a site of injury, the formation of blood clots BRIEF SUMMARY OF THE INVENTION at inappropriate Sites in the body can also produce harmful 0009. The present invention includes isolated nucleic effects. Examples of particularly dangerous blood clots acid molecules comprising, or alternatively, consisting of a include embolisms resulting from free floating blood clots polynucleotide Sequence disclosed in the Sequence listing which have become detatched from their site of formation and/or contained in a human cDNA plasmid described in elsewhere in the circulatory System. Embolisms arising from Table I and deposited with the American Type Culture free-floating blood clots may result in Such pathologies as Collection (ATCC). Fragments, variants, and derivatives of Strokes and heart attacks. Furthermore, blood clots may these nucleic acid molecules are also encompassed by the become lodged in Smaller channels of the circulatory SyS invention. The present invention also includes isolated tem, resulting in Stenosis, or narrowing, of the circulatory nucleic acid molecules comprising, or alternatively, consist System passage. Stenosis may diminish the flow of blood to ing of, a polynucleotide encoding plasminogen-like particular organs or regions of the body. polypeptides. The present invention further includes plas 0005 The natural process opposing blood clot formation minogen-like polypeptides encoded by these polynucle is fibrinolysis, in which the blood clot is solublized to restore otides. Further provided for are amino acid Sequences com normal blood flow. The principal mediator of fibrinolysis is prising, or alternatively, consisting of, plasminogen-like US 2003/0180934 A1 Sep. 25, 2003 polypeptides as disclosed in the Sequence listing and/or 0017. In the present invention, “isolated” refers to mate encoded by the human cDNA plasmids described in Table 1 rial removed from its original environment (e.g., the natural and deposited with the ATCC. Antibodies that bind these environment if it is naturally occurring), and thus is altered polypeptides are also encompassed by the invention. “by the hand of man” from its natural state. For example, an Polypeptide fragments, variants, and derivatives of these isolated polynucleotide could be part of a vector or a amino acid Sequences are also encompassed by the inven composition of matter, or could be contained within a cell, tion, as are polynucleotides encoding these polypeptides and and still be “isolated” because that vector, composition of antibodies that bind these polypeptides. matter, or particular cell is not the original environment of the polynucleotide. The term "isolated” does not refer to DETAILED DESCRIPTION OF THE genomic or cDNA libraries, whole cell total or mRNA INVENTION preparations, genomic DNA preparations (including those Separated by electrophoresis and transferred onto blots), 0010 Tables Sheared whole cell genomic DNA preparations or other 0.011 Table 1 summarizes ATCC Deposits, Deposit dates, compositions where the art demonstrates no distinguishing and ATCC designation numbers of deposits made with the features of the polynucleotide/Sequences of the present ATCC in connection with the present
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