Erythropoietin: remodeling and glycoconjugation of erythropoietin

Abstract

The invention includes methods and compositions for remodeling a peptide molecule, including the addition or deletion of one or more glycosyl groups to a peptide, and / or the addition of a modifying group to a peptide.

Description

CROSS REFERENCE TO RELATED APPLICATIONS [0001] This application is a divisional application of U.S. patent application Ser. No. 10 / 410,945, filed Apr. 9, 2003; which is a continuation-in-part of prior International Application No. PCT / US02 / 32263, filed Oct. 9, 2002 and published in English under Article 21(2) of the PCT as WO. This application claims the benefit under 35 USC 119(e) of U.S. Provisional Patent Application No. 60 / 407,527, filed Aug. 28, 2002; U.S. Provisional Patent Application No. 60 / 404,249, filed Aug. 16, 2002; U.S. Provisional Patent Application No. 60 / 396,594, filed Jul. 17, 2002; U.S. Provisional Patent Application No. 60 / 391,777, filed Jun. 25, 2002; U.S. Provisional Patent Application No. 60 / 387,292, filed Jun. 7, 2002; U.S. Provisional Patent Application No. 60 / 334,301, filed Nov. 28, 2001; U.S. Provisional Patent Application No. 60 / 334,233, filed Nov. 28, 2001; U.S. Provisional Patent Application No. 60 / 344,692, filed Oct. 19, 2001; and U.S. Provisional Paten...

AI Technical Summary

Benefits of technology

[0184] Further provided is a method of treating a kidney dialysis patient, the method comprising administering to the patient a glycoPEGylated EPO peptide comprising an EPO peptide and at least one glycan and at least one poly(ethylene glycol) molecule covalently attached to the glycan, wherein the poly(ethylene glycol) molecule is added to the EPO using a glycosyltransferase, wherein the EPO is administered in an amount effective to increase the hematocrit level in the patient.

Problems solved by technology

The alteration of one or more of these characteristics greatly affects the efficacy of a peptide in its natural setting, and also affects the efficacy of the peptide as a therapeutic agent in situations where the peptide has been generated for that purpose.

Peptides whose glycans do not contain terminal sialic acid residues are rapidly removed from the circulation by the liver, an event which negates any potential therapeutic benefit of the peptide.

A major factor, which has limited the use of therapeutic peptides is the immunogenic nature of most peptides.

Other deficiencies of therapeutic peptides include suboptimal potency and rapid clearance rates.

The tetrasaccharide sialyl Lewis X was then enzymatically rebuilt on the remaining GlcNAc anchor site on the now homogenous protein by the sequential use of β-1,4-galactosyltransferase, α-2,3-sialyltransferase and α-1,3-fucosyltransferase V. However, while each enzymatically catalyzed step proceeded in excellent yield, such procedures have not been adapted for the generation of glycopeptides on an industrial scale.

Further, the methods of cell surface modification are not utilized for the enzymatic incorporation preformed modified glycosyl donor moiety into a peptide.

Moreover, none of the cell surface modification methods are practical for producing glycosyl-modified peptides on an industrial scale.

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