Use of Polysaccharides for Promotion of Enzymatic Activity

Abstract

This disclosure provides methods and compositions for the promotion of enzymatic activity of Target Enzymes, including but not limited to oligosaccharide/polysaccharide enzymes, protein enzymes, polynucleotide enzymes. The methods involve use of a non-naturally occurring polysaccharide (including but not limited HES) for promoting the enzymatic activity of an enzyme in liquid milieu, wherein the concentration of the polysaccharide in the composition comprising the Target Enzyme is from about 0.01% to about 55% w/v.

1 . A composition comprising:
 a) a Target Enzyme selected from the group consisting of: an oligosaccharide/polysaccharide enzyme, a protein enzyme, a polynucleotide enzyme, and a lipase, and;   b) a non-naturally occurring starch,
 wherein the composition comprises from about 0.01% to about 55% w/v of the polysaccharide and wherein the activity of the Target Enzyme has been promoted by the non-naturally occurring polysaccharide. 
   
     
     
         2 . The composition of  claim 1 , wherein the Target Enzyme is
 an oligosaccharide/polysaccharide enzyme selected from the group consisting of: Galactosyltransferase, GaINAc transferase, Oligosaccharyltransferase, N-acetylglucosaminylphosphotransferase, O-linked glycosyltransferase, N-linked glycosyltransferase, α-Mannosidase, β-Galactosidase, Sialidase (neuraminidase), β-N-acetylhexosaminidase, N-Acetyl-glucosamine-1-phosphodiester α-N-acetylglucosaminidase, N-Glycanase (N-glycosidase F), O-Glycanase (endo-α-N-acetylgalactosaminidase), Endo-β-N-acetylglucosaminidase H, Sialate-O-acetyltransferase, Sialate-O-acetylesterase, and Alpha-glucosidase;   a protein enzyme(s) selected from the group consisting of: Protease, Myristoylase, Deformylase, N-Terminal methionine excision enzyme, Phosphorylase, Acetylation enzyme, Disulfide bond formation enzyme, Palmitoylation enzyme, Hydroxylation enzyme, Carboxylation enzyme, Nitration enzyme, Sulfation enzyme, ADP-ribosylation enzyme, Deamidase, N-Linked glycosylase, O-Linked glycosylase, Glycosyl-phosphoinositolation enzyme, Farnesylase, Geranylgeranylase, Methylase, Amidation enzyme, and Ubiquitination enzyme;   a polynucleotide enzyme selected from the group consisting of: Exoribonuclease, Endoribonuclease, Exodeoxyribonuclease, Endodeoxyribonuclease, Restriction endonuclease, Topoisomerase I, Topoisomerase II, and Ligase; or   a lysosomal hydrolase selected from the group consisting of: α-Galactosidase A, Acid ceramidase, Acid α-L-fucosidase, Acid/β-glucocerebrosidase (GCR), Acid β-galactosidase, lduronate-2-sulfatase, α-L-lduronidase, Galactocerebrosidase, Acid α-mannosidase, Acid β-mannosidase, Arylsulfatase B, Arylsulfatase A, N-Acetylgalactosamine-6-sulfate sulfatase, Acid β-galactosidase, Acid sphingomyelinase, Acid α-glucosidase, β-Hexosaminidase B, Heparan N-sulfatase, α-N-Acetylglucosaminidase, Acetyl-CoA: α-glucosaminide N-acetyltransferase, N-Acetylglucosamine-6-sulfate sulfatase, α-N-acetylgalactosaminidase, Sialidase, β-Glucuronidase, and β-Hexosaminidase A.   
     
     
         3 . The composition of  claim 1 , wherein the non-naturally occurring starch is a hydroxyalkyl starch. 
     
     
         4 . The composition of  claim 3 , wherein the hydroxyalkyl starch is HES. 
     
     
         5 . A composition comprising:
 a) a Target Enzyme selected from the group consisting of:
 an oligosaccharide/polysaccharide enzyme, a protein enzyme, a polynucleotide enzyme, and a lipase, and; 
   b) a non-naturally occurring starch,
 wherein the composition comprises from about 1% to about 50% w/v of the polysaccharide. 
   
     
     
         6 . The composition of  claim 5  wherein the composition comprises from about 10% to about 40% w/v of the polysaccharide. 
     
     
         7 . A pharmaceutical composition comprising:
 a) a Target Enzyme which is a therapeutic protein and;   b) a non-naturally occurring starch,
 wherein the composition comprises from about 0.01% to about 55% w/v of the polysaccharide. 
   
     
     
         8 . A composition according to  claim 1 , further comprising a substrate of the Target Enzyme. 
     
     
         9 . The composition of  claim 8 , wherein the substrate is selected from the group consisting of a protein, a peptide, a polynucleotide, a nucleotide, and a small molecule. 
     
     
         10 . The composition of  claim 9 , wherein the substrate is an enzyme. 
     
     
         11 . The composition of  claim 10 , wherein the substrate is a lysosomal hydrolase. 
     
     
         12 . The composition of  claim 11 , wherein the lysosomal hydrolase is Acid/β-glucocerebrosidase (GCR). 
     
     
         13 . The composition of  claim 12  wherein the Target Enzyme is selected from the group consisting of: sialidase, β-galactosidase, β-N-acetylhexosaminidase and a combination thereof; the non-naturally occurring polysaccharide is HES; and the substrate is β-glucocerebrosidase. 
     
     
         14 . A composition comprising:
 a) an oligosaccharide/polysaccharide enzyme;   b) between about 1% and about 12% w/v HES; and   c) a substrate for the oligosaccharide/polysaccharide enzyme.   
     
     
         15 . The composition of  claim 14 , wherein the oligosaccharide/polysaccharide enzyme is selected from the group consisting of: sialidase, β-galactosidase, β-N-acetylhexosaminidase and a combination thereof. 
     
     
         16 . The composition of  claim 14 , further comprising citrate buffer and calcium chloride at a pH of about 6. 
     
     
         17 - 30 . (canceled)