US2005214276A9PendingUtilityA9

Delta 4, 5 glycuronidase and uses thereof

53
Individually held — no corporate assignee on recordPriority: May 3, 2002Filed: May 5, 2003Published: Sep 29, 2005
Est. expiryMay 3, 2022(expired)· nominal 20-yr term from priority
Inventors:James R. MyetteZachary ShriverGanesh VenkataramanRam SasisekharanMaitland W. Mclean
C07H 21/04A61P 7/00A61P 9/00A61P 43/00A61K 38/00C12Y 302/01166C12Y 302/01056C12Q 1/34A61P 35/00A61P 7/04C12N 9/2402Y02A50/30
53
PatentIndex Score
0
Cited by
0
References
0
Claims

Abstract

The invention relates to Δ4,5 glycuronidase, related compositions, and methods of use thereof.

Claims

exact text as granted — not AI-modified
1 . A substantially pure Δ4,5 unsaturated glycuronidase.  
     
     
         2 . The glycuronidase of  claim 1 , wherein the glycuronidase is a recombinantly produced glycuronidase.  
     
     
         3 . The glycuronidase of  claim 1 , wherein the glycuronidase is a synthetic glycuronidase.  
     
     
         4 . The glycuronidase of  claim 1 , wherein the glycuronidase is a peptide having an amino acid sequence of SEQ ID NO:1, or a functional variant thereof.  
     
     
         5 . An isolated polypeptide comprising: 
 Δ4,5 unsaturated glycuronidase having an amino acid sequence of SEQ ID NO: 1 or a functional variant thereof.    
     
     
         6 . The isolated polypeptide of  claim 5 , wherein the glycuronidase has the amino acid sequence of SEQ ID NO: 1.  
     
     
         7 . A composition comprising, an isolated Δ4,5 unsaturated glycuronidase having a higher specific activity than native glycuronidase.  
     
     
         8 . The composition of  claim 7 , wherein the glycuronidase has a specific activity of at least about 60 picomoles of substrate hydrolyzed per minute per picomole of glycuronidase.  
     
     
         9 . The composition of  claim 7 , wherein the Δ4,5 unsaturated glycuronidase has at least about a two fold higher specific activity than native glycuronidase.  
     
     
         10 . The composition of  claim 7 , wherein the Δ4,5 unsaturated glycuronidase has at least about a three fold higher specific activity than native glycuronidase.  
     
     
         11 . The composition of  claim 7 , wherein the Δ4,5 unsaturated glycuronidase has at least about a four fold higher specific activity than native glycuronidase.  
     
     
         12 . An isolated nucleic acid molecule selected from the group consisting of 
 (a) nucleic acid molecules which hybridize under stringent conditions to a nucleic acid molecule having a nucleotide sequence set forth as SEQ ID NO:2, and which code for Δ4,5 unsaturated glycuronidase having an amino acid sequence set forth as SEQ ID NO:1,    (b) nucleic acid molecules that differ from the nucleic acid molecules of (a) in codon sequence due to degeneracy of the genetic code, and    (c) complements of (a) or (b).    
     
     
         13 . The isolated nucleic acid molecule of  claim 12 , wherein the isolated nucleic acid molecule codes for SEQ ID NO:1.  
     
     
         14 . The isolated nucleic acid molecule of  claim 12 , wherein the isolated nucleic acid molecule comprises the nucleotide sequence set forth as SEQ ID NO:2.  
     
     
         15 . An expression vector comprising the isolated nucleic acid molecule of  claim 10  operably linked to a promoter.  
     
     
         16 . A host cell comprising the expression vector of  claim 15 .  
     
     
         17 . An expression vector comprising the isolated nucleic acid molecule of  claim 14  operably linked to a promoter.  
     
     
         18 . A host cell comprising the expression vector of  claim 17 .  
     
     
         19 . A pharmaceutical preparation comprising: 
 a composition or vector of any of claims  1 - 15  and a pharmaceutically acceptable carrier.    
     
     
         20 . A method of cleaving a glycosaminoglycan, comprising: 
 contacting a glycosaminoglycan with the glycuronidase of any one of claims  1 - 10  in an effective amount to cleave the glycosaminoglycan.    
     
     
         21 . A glycosaminoglycan prepared according to the method of  claim 20 .  
     
     
         22 . A method of removing heparin from a heparin containing fluid, comprising: 
 contacting a heparin containing fluid with the glycuronidase of any one of claims  1 - 10  in an effective amount to removing heparin from the heparin containing fluid.    
     
     
         23 . The method of  claim 22 , wherein the glycuronidase is immobilized on a solid support.  
     
     
         24 . The method of  claim 23 , wherein heparinase is also immobilized on the solid support.  
     
     
         25 . A method of analyzing of a glycosaminoglycan, comprising: 
 contacting a glycosaminoglycan with the glycuronidase of any one of claims  1 - 10  in an effective amount to analyze the glycosaminoglycan.    
     
     
         26 . The method of  claim 25 , wherein the method is a method for identifying the presence of a particular glycosaminoglycan in a sample.  
     
     
         27 . The method of  claim 25 , wherein the method is a method for determining the purity of a glycosaminoglycan in a sample.  
     
     
         28 . The method of  claim 25 , wherein the method is a method for determining the composition of a glycosaminoglycan in a sample.  
     
     
         29 . The method of  claim 25 , wherein the method is a method for determining the sequence of saccharide units in a glycosaminoglycan.  
     
     
         30 . The method of  claim 29 , further comprising an additional analytic technique selected from the group consisting of mass spectrometry, NMR spectroscopy, gel electrophoresis, capillary electrophoresis, and HPLC.  
     
     
         31 . A method of cleaving a glycosaminoglycan comprised of at least one disaccharide unit, comprising: 
 contacting a glycosaminoglycan comprised of at least one disaccharide unit with the glycuronidase of any one of claims  1 - 10  in an effective amount to cleave the glycosaminoglycan.    
     
     
         32 . The method of  claim 31 , wherein the glycosaminoglycan is comprised of ΔUH NAc  disaccharide units.  
     
     
         33 . The method of  claim 31 , wherein the glycosaminoglycan is comprised ΔUH NAc,6S  disaccharide units.  
     
     
         34 . The method of  claim 31 , wherein the glycosaminoglycan is comprised of ΔUH NS,6S  disaccharide units.  
     
     
         35 . The method of  claim 31 , wherein the glycosaminoglycan is comprised of ΔUH NS  disaccharide units.  
     
     
         36 . The method of  claim 31 , wherein the glycosaminoglycan length exceeds two saccharide units.  
     
     
         37 . The method of  claim 30 , wherein the glycosaminoglycan does not contain a 2-O sulfated uronidate.  
     
     
         38 . The method of  claim 30 , wherein the glycosaminoglycan is 6-0 sulfated.  
     
     
         39 . The method of  claim 30 , wherein the glycosaminoglycan does contain an N-unsubstituted glycosamine.  
     
     
         40 . A method of inhibiting angiogenesis, comprising 
 administering to a subject in need thereof an effective amount of the pharmaceutical preparation of  claim 19  for inhibiting angiogenesis.    
     
     
         41 . A method of treating cancer, comprising 
 administering to a subject in need thereof an effective amount of the pharmaceutical preparation of  claim 19  for treating cancer.    
     
     
         42 . A method of inhibiting cellular proliferation, comprising 
 administering to a subject in need thereof an effective amount of the pharmaceutical preparation of  claim 19  for of inhibiting cellular proliferation.    
     
     
         43 . A method of treating a coagulation disorder, comprising 
 administering the composition of  claim 19  to a subject in need thereof an effective amount for treat the coagulation disorder.    
     
     
         44 . The method of  claim 20 , wherein the use of the glycuronidase is concurrent with or follows treatment with heparinase.  
     
     
         45 . A pharmaceutical preparation comprising: 
 a composition of  claim 19  and a pharmaceutically acceptable carrier.    
     
     
         46 . A method of inhibiting angiogenesis, comprising 
 administering the composition of  claim 21  to a subject in need thereof an effective amount for inhibiting angiogenesis.    
     
     
         47 . A method of treating cancer, comprising 
 administering the composition of  claim 21  to a subject in need thereof an effective amount for treating cancer.    
     
     
         48 . A method of inhibiting cellular proliferation, comprising 
 administering the composition of  claim 21  to a subject in need thereof an effective amount for inhibiting cellular proliferation.    
     
     
         49 . A method of treating a coagulation disorder, comprising 
 administering the composition of  claim 21  to a subject in need thereof an effective amount for treat the coagulation disorder.

Join the waitlist — get patent alerts

Track US2005214276A9 — get alerts on status changes and closely related new filings.

We store only your email — no account needed. See our privacy policy.