Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Rodent cell – per se
Reexamination Certificate
2000-10-10
2003-04-01
Monshipouri, M. (Department: 1652)
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Rodent cell, per se
C435S366000, C435S371000, C435S196000, C536S023200
Reexamination Certificate
active
06541254
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to glycosylation variants of iduronate-2-sulfatase and to genetic sequences encoding same. The present invention also contemplates the use of these in the treatment and diagnosis of subjects suspected of, or suffering from, iduronate 2-sulfatase deficiency.
BACKGROUND TO INVENTION
Iduronate 2-sulfatase (hereinafter abbreviated to “IDS”; EC 3.1.6.13) acts as an exosulfatase in lysosomes to hydrolyze the C2-sulfate ester bond from non-reducing-terminal iduronic acid residues in the glycosaminoglycans heparan sulfate and dermatan sulfate (1). IDS is one of a family of at least nine sulfatases that hydrolyze sulfate esters in human cells. They are all lysosomal enzymes that act on sulfated monosaccharide residues in a variety of complex substrates with the exception of microsomal steroid sulfatase (or arylsulfatase C), which acts on sulfated 3&agr;-hydroxysteriods (1,2). Each sulfatase displays absolute substrate specificity, making the sulfatase family an attractive model to investigate the molecular requirements for substrate binding and the catalysis of sulfate ester hydrolysis.
A deficiency in the activity of IDS in humans leads to the lysosomal accumulation of heparan sulfate and dermatan sulfate fragments and their excretion in urine (1). This storage results in the clinical disorder Hunter syndrome (mucopolysaccharidosis type II, MPS-II) in which patients may present with variable phenotypes from severe mental retardation, skeletal deformities, and stiff joints to a relatively mild course (1). It has been postulated that this clinical heterogeneity reflects different mutations at the IDS locus affecting enzyme expression, stability, or function. MPS-II is one of the most common mucopolysaccharidoses and is the only one that is X chromosome-linked (1).
In accordance with the present invention, there is provided the nucleotide sequence for a full length cDNA clone for IDS from human endothelial cells. The present invention also provides the genomic clone for IDS. More particularly, following expression of the IDS nucleotide sequence in particular cell lines, a glycosylation variant of IDS has been isolated which possesses inter alia improved half-life and/or improved uptake properties when compared to the naturally glycosylated molecule.
SUMMARY OF THE INVENTION
One aspect of the present invention provides a recombinant human iduronate 2-sulfatase (IDS) or a fragment thereof retaining enzymatic activity wherein said recombinant IDS or fragment thereof is more highly glycosylated than the naturally occurring enzyme or equivalent fragment on the naturally occurring enzyme.
Another aspect of the present invention contemplates a method for treating a patient suffering from iduronate 2-sulfatase (IDS) deficiency said method comprising administering to said patient an effective amount of a recombinant human IDS or a fragment thereof retaining enzymatic activity wherein said recombinant IDS or fragment thereof is more highly glycosylated than the naturally occuring enzyme or equivalent fragment on the naturally occurring enzyme.
Yet another aspect of the present invention is directed to a pharmaceutical composition useful in the treatment of patients suffering from iduronate 2-sulfatase (IDS) deficiency said composition comprising the more highly glycosylated IDS or enzymatically active fragment thereof referred to above and one or more pharmaceutically acceptable carriers and/or diluents.
Still yet another aspect of the present invention provides an isolated genomic DNA fragment carrying in whole or in part the IDS gene or a mutant or derivative thereof. The isolation of the genomic clone will enable gene therapy and genetic analysis of IDS deficiency diseases.
REFERENCES:
patent: 4837147 (1989-06-01), Oshima et al.
Anson, et al. (1992) “Correction of human mucopolysaccharidosis type VI fibroblasts with recombinant N-acetylgalactosamine-4-sulphatase”,Biochem J. 284: 789-794.
Bielicki, et al. (1990) “Human liver iduronate-2-sulphatase”,Biochem J. 271: 75-86.
Bielicki, et al. (1993) “Recombinant Human Iduronate-2-Sulphatase: Correction of Mucopoly-saccharidosis—Type II Fibroblasts and Characterization of The Purified Enzyme”,J. Biochem. 289: 241-246.
Darnell, et al. (1986) “Protein Glycosylation”,Molecular Cell Biology: 957-964.
Ferrara, et al. (1987) “Characterization of recombinant glycosylated human interleukin 2 produced by a recombinant plasmid transformed CHO cell line”,FEBS Letters 226(1); 47-52.
Gottesman, Michael M. (1987) “Chinese Hamster Ovary Cells”,Methods in Enzymology 151: 3-8.
Kodama, et al. (1992) “Characterization of recombinant murine interleukin 5 expressed in Chinese hamster ovary cells”,Glycobiology 2(5): 419-427.
Peters, et al. (1990) “Phylogenetic Conservation of Arylsulfatases”,J. Biol. Chem. 265(6): 3374-3381.
Robertson, et al. (1988) “Human Glucosamine-6-sulfatase cDNA Reveals Homology with Steroid Sulfatase”,Biochem. Biophys. Res. Commun. 157(1): 218-224.
Sasaki, et al. (1988) “cDNA cloning, nucleotide sequence and expression of the gene for arylsulfatase in the sea urchin (Hemicentrous pulcherrimus) embryo”,Eur. J. Biochem. 177: 9-13.
Stein, et al. (1989) “Cloning and Expression of Human Arylsulfatase A”,J. Biol. Chem. 264(2): 1252-1259.
Stein, et al. (1989) “Cloning and Expression of Human Steroid-sulfatase”,J. Biol. Chem. 264(23): 13865-13872.
Wilson, et al. (1990) “Hunter Syndrome: Isolation of an Iduronate-2-Sulfatase cDNA clone and Analysis of Patient DNA”,Proc. Natl. Acad. Sci. USA 87: 8531-8535.
Yen, et al. (1987) “Cloning and Expression of Steroid Sulfatase cDNA and the Frequent Occurence of Deletions in STS Deficiency: Implications for X-Y Interchange”,Cell 49: 443-454.
Yen, et al. (1988) “The Human X-Linked Steroid Sulfatase Gene and a Y-Encoded Pseudogene: Evidence for an Inversion of the Y Chromosome during Primate Evolution”,Cell 55: 1123-1135.
Anson Donald Stewart
Bielicki Julie
Clements Peter Roy
Hopwood John Joseph
Morris Charles Phillip
Dilworth & Barrese LLP
Monshipouri M.
Pokalsky Ann R.
Women's and Children's Hospital
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