SACCHARIDE-CONTAINING PROTEIN CONJUGATES AND USES THEREOF
Conjugates of a saccharide and a biomolecule, covalently linked therebetween via a non-hydrophobic linker and methods of preparing same are disclosed. Also disclosed are medical uses utilizing such conjugates. Glycosylation reagents for use in preparing these conjugates are also disclosed. Glycosylated proteins, characterized by improved performance, are also disclosed.
1 . A compound comprising a saccharide moiety and a non-hydrophobic linker being attached thereto, said linker comprising a poly(alkylene glycol) chain of at least 18 atoms in length, said poly(alkylene glycol) comprising 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 alkylene glycol units.
2 . The compound of claim 1 , wherein said linker is attached to an anomeric carbon of said saccharide moiety.
3 . The compound of claim 2 , wherein said linker is attached to said anomeric carbon via a bond having α configuration.
4 . The compound of claim 1 , wherein said poly(alkylene glycol) comprises poly(ethylene glycol) (PEG).
5 . The compound of claim 1 , wherein said poly(alkylene glycol) is from 24 to 36 atoms in length.
6 . The compound of claim 4 , wherein said poly(ethylene glycol) comprises from 8 to 12 ethylene glycol units.
7 . The compound of claim 1 , wherein said linker is a branched linker comprising at least two linear chemical moieties which are covalently linked to one another, each of said linear chemical moieties being attached to a saccharide moiety, such that said linker is attached to at least two saccharide moieties.
8 . The compound of claim 7 , wherein said branched linker comprises at least two linear poly(alkylene glycol) moieties which are covalently linked to one another.
9 . The compound of claim 1 , wherein said saccharide moiety is a monosaccharide, and said linker is a non-saccharide linker.
10 . The compound of claim 9 , wherein said monosaccharide is selected from the group consisting of a sialic acid, a mannose and a M6P.
11 . The compound of claim 1 , wherein said saccharide moiety is a hexose.
12 . The compound of claim 11 , wherein said hexose is a D-hexose.
13 . The compound of claim 1 , wherein said linker comprises a single reactive group.
14 . The compound of claim 13 , wherein said reactive group is capable of reacting with a protein to form a covalent bond between said linker and said protein.
15 . The compound of claim 13 , wherein said reactive group is selected from the group consisting of an amine, a maleimide and a carboxylate.
16 . The compound of claim 1 having the formula:
wherein:
n=8; and
Z is selected from the group consisting of —CH 2 CH 2 CO 2 H and —CH 2 CH 2 —NHCOCH 2 CH 2 -maleimide.
17 . A compound selected from the group consisting of:
18 . A compound comprising a monosaccharide moiety and a non-hydrophobic linker being attached thereto, said monosaccharide being selected from the group consisting of sialic acid and M6P and said linker being a non-saccharide linker and comprising a poly(alkylene glycol) being at least 18 atoms in length.
19 . The compound of claim 18 , wherein said linker comprises a single reactive group.
20 . A conjugate formed by reacting the compound of claim 14 with a protein, to thereby form said covalent bond between said linker and said protein.