Glycopegylated erythropoietin
AssigneeNeose Technologies, Inc.
InventorsShawn DeFrees, Robert J. Bayer, David A. Zopf
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Abstract
The present invention provides conjugates between erythropoietin and PEG moieties. The conjugates are linked via an intact glycosyl linking group interposed between and covalently attached to the peptide and the modifying group. The conjugates are formed from glycosylated peptides by the action of a glycosyltransferase. The glycosyltransferase ligates a modified sugar moiety onto a glycosyl residue on the peptide. Also provided are methods for preparing the conjugates, methods for treating various disease conditions with the conjugates, and pharmaceutical formulations including the conjugates.
Claims36
An erythropoietin peptide conjugate comprising a sialic acid moiety having the formula:
- wherein
- D is a member selected from —OH and R1-L-HN—;
- G is a member selected from R1-L- and —C(O)(C1-C6)alkyl;
- R1 is a moiety comprising a branched poly(ethylene glycol) residue;
- and
- L is a linker which is a member selected from a bond, substituted or
- unsubstituted alkyl and substituted or unsubstituted heteroalkyl,
- such that when D is OH, G is R1-L-, and when G is —C(O)(C1-C6)alkyl, D is R1-L-NH—,
- wherein said branched poly(ethylene glycol) residue has a formula which is a member selected from:
- wherein
- q1 is an integer from 0 to 20;
- q, q′ and q″ are integers independently selected from from 1 to 20;
- and
- e, f and f′ are integers independently selected 1 to 2500.
A method of making a PEG-ylated erythropoietin comprising the moiety:
- wherein
- D is a member selected from -OH and R1-L-NH-;
- G is a member selected from R1-L- and -C(O)(C1-C6)alkyl;
- R1 is a moiety comprising a straight-chain or branched poly(ethylene glycol) residue;
- and
- L is a linker which is a member selected from a bond, substituted or unsubstituted alkyl and substituted or unsubstituted heteroalkyl,
- such that when D is OH, G is R1-L-, and when G is -C(O)(C1-C6)alkyl, D is R1-L-NH-,
- said method comprising:
- (a) contacting a substrate erythropoietin peptide comprising the glycosyl moiety:
- with a PEG-sialic acid donor moiety having the formula:
- and an enzyme that transfers said PEG-sialic acid onto the Gal of said glycosyl moiety, under conditions appropriate to for said transfer.
Dependencies are inferred from the claim text.