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  Chondroitin sulfate synthesis promoterUSPTO Application #: 20070203087Title: Chondroitin sulfate synthesis promoter Abstract: The invention provides a chondroitin sulfate synthesis promoter useful for the treatment of diseases such as articular disease and discopathy. The chondroitin sulfate synthesis promoter contains, as an active ingredient, chondroitin sulfate glucuronyltransferase protein and/or chondroitin sulfate N-acetylgalactosaminyltransferase-1 protein, or a gene encoding the enzyme protein(s). (end of abstract) Agent: Factor & Lake, Ltd - Chicago, IL, US Inventors: Kenichiro SAKAI, Koji KIMATA, Kenichi SHINOMIYA, Hideto WATANABE USPTO Applicaton #: 20070203087 - Class: 514 44 (USPTO) The Patent Description & Claims data below is from USPTO Patent Application 20070203087. Brief Patent Description - Full Patent Description - Patent Application Claims
CROSS-REFERENCE TO RELATED APPLICATIONS [0001]This application claims priority to Japanese application number 2006-045813, filed on Feb. 22, 2006 and Japanese application number 2006-224252, filed on Aug. 21, 2006, both of which are incorporated herein by reference. BACKGROUND OF THE INVENTION [0002]1. Field of the Invention [0003]The present invention relates to a chondroitin sulfate synthesis promoter, and to a treatment agent for articular disease and discopathy employing the promoter. [0004]2. Background Art [0005]Cartilage contains a macromolecular substance, called aggrecan, which is predominantly formed of chondroitin sulfate proteoglycan. Aggrecan is a type of proteoglycan in which a polysaccharide sulfate is covalently bound to a core protein, and is known to play an important role in organisms. In cartilage, aggrecan, having a hyaluronic acid-binding domain in a core protein portion, forms a macromolecular complex with a link protein and hyaluronic acid. Aggrecan, having 100 or more chondroitin sulfate chains bound to a core protein portion thereof, retains a large number of water molecules. Thus, aggrecan is present in cartilage in the form of highly hydrated gel and is believed to play important roles such as absorbing shock and reducing friction (lubrication). [0006]Meanwhile, several glycosyltransferases have recently been reported to be involved in synthesis of chondroitin sulfate chains: chondroitin synthase-1 (CSS-1, J. Biol. Chem., October 2001; 276: 38721-38726), chondroitin synthase-2 (CSS-2, J. Biol. Chem., August 2003; 278: 30235-30247, WO 03/102194), chondroitin synthase-3 (CSS-3, J. Biol. Chem., October 2003; 278: 39711-39725, WO 03/102193), chondroitin sulfate glucuronyltransferase (CSGlcAT, J. Biol. Chem., October 2002; 277: No. 41: 38179-38188), chondroitin sulfate N-acetylgalactosaminyltransferase-1 (CSGalNAcT-1, J. Biol. Chem., March 2002; 277: No. 11: 8841-8846), and chondroitin sulfate N-acetylgalactosaminyltransferase-2 (CSGalNAcT-2, J. Biol. Chem., January 2003; 278: 3063-3071, Japanese Patent Application Laid-Open (kokai) No. 2003-289883). [0007]Although general functions of these enzymes are already known, the roles of these enzymes in organisms have not yet been elucidated. In addition, an enzyme involved in biosynthesis of chondroitin sulfate (a part of aggrecan biosynthesis) has not been identified. Therefore, whether or not these enzymes increase the amount of chondroitin sulfate in organisms remains unknown. SUMMARY OF THE INVENTION [0008]Degradation of cartilage is accelerated by aging or overload. For example, in recent aging society, an increased number of elderly people suffer knee joint pain caused by wear of knee cartilage, leading to keen demand for means of repairing cartilage functions. In cartilage of elderly people, shock absorption performance and friction reduction performance (lubricating action) are considerably impaired. Loss of functions intrinsic to cartilage may predominantly be attributable to a decrease in the amount of chondroitin sulfate chains contained by proteoglycan such as aggrecan. Under such circumstances, the present inventors have attempted to repair and improve functions of cartilage through promoting biosynthesis of chondroitin sulfate chains of aggrecan. [0009]In order to solve the aforementioned object, the present inventors have conducted extensive studies, and have found that among chondroitin sulfate synthesis-related enzymes, chondroitin sulfate glucuronyltransferase (hereinafter sometimes abbreviated as CSGlcAT) and chondroitin sulfate N-acetylgalactosaminyltransferase-1 (hereinafter sometimes referred to as CSGalNAcT-1) are expressed in mouse embryonic cartilage and mouse cartilage differentiated cells in an expression pattern which is considerably correlated with a known aggrecan expression pattern. The inventors have also found that forced over-expression of CSGlcAT and CSGalNAcT-1 in chondroid cells remarkably promotes synthesis of chondroitin sulfate. The inventors have further found that introduction of CSGalNAcT-1 Gene to mouse intervertebral discs increases the levels of chondroitin sulfate in the intervertebral discs, through in vivo experiment. [0010]The present invention has been accomplished on the basis of these findings. [0011]Accordingly, in one aspect of the present invention, there is provided a chondroitin sulfate synthesis promoter comprising, as an active ingredient, a gene encoding chondroitin sulfate glucuronyltransferase and/or a gene encoding chondroitin sulfate N-acetylgalactosaminyltransferase-1. [0012]Preferably, the gene encoding chondroitin sulfate glucuronyltransferase is a nucleic acid molecule selected from at least one of the following: [0013](a): a nucleic acid molecule including a nucleotide sequence defined by SEQ ID NO: 1, [0014](b): a nucleic acid molecule which can be hybridized with a complement of a nucleic acid of SEQ ID NO: 1 under stringent conditions, or [0015](c): a nucleic acid molecule including a nucleotide sequence identical to that defined by SEQ ID NO: 1, except that one or more nucleotide(s) have been substituted, deleted, inserted, or transposed, wherein the nucleic acid molecule encodes a protein having activity of transferring glucuronate from a glucuronate donor to a non-reducing end N-acetylgalactosamine residue present in the chondroitin skeleton. [0016]Preferably, the chondroitin sulfate synthesis promoter comprises, as an active ingredient, a nucleic acid molecule including a nucleotide sequence encoding an amino acid sequence defined by SEQ ID NO: 2. [0017]Preferably, the gene encoding chondroitin sulfate N-acetylgalactosaminyltransferase-1 is a nucleic acid molecule selected from at least one of the following: [0018](d): a nucleic acid molecule including a nucleotide sequence defined by SEQ ID NO: 3, [0019](e): a nucleic acid molecule which can be hybridized with a complement of a nucleic acid of SEQ ID NO: 3 under stringent conditions, or [0020](f): a nucleic acid molecule including a nucleotide sequence identical to that defined by SEQ ID NO: 3, except that one or more nucleotide(s) have been substituted, deleted, inserted, or transposed, wherein the nucleic acid molecule encodes a protein having activity of transferring an N-acetylgalactosamine residue from an N-acetylgalactosamine donor to a non-reducing end D-glucuronate residue of an N-acetylgalactosamine acceptor substrate containing a sugar chain represented by formula; GlcUA-Gal-Gal-Xyl Continue reading... 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