Method of preparing a modified granulocyte colony stimulating factor (G-CSF) with reduced immunogenicity

US7392141B2 · kind B2 · utility

Assignee

• Merck Patent GmbH · DE

Inventors

• Francis Joseph Carr · Aberdeen, GB
• Graham Carter · Dyce, GB
• Tim Jones · Cary, US
• Stephen Williams · Laramie, US

Key dates

• Filing date: Feb 5, 2002
• Grant date: Jun 24, 2008
• Expiry date: Dec 29, 2023

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10S707/99943
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

A method of preparing a modified granulocyte colony stimulating factor (G-CSF) protein having reduced immunogenicity relative to human G-CSF comprises the steps of (i) identifying one or more potential T-cell epitopes within the amino acid sequence of human G-CSF (SEQ ID NO: 1); (ii) designing at least one sequence variant of at least one potential T-cell epitope identified in step (i), wherein the sequence variant eliminates or substantially reduces the MHC class II binding activity of the potential T-cell epitope; (iii) preparing, by recombinant DNA techniques, at least one modified G-CSF protein including a sequence variant designed in step (ii); (iv) evaluating at least one modified G-CSF protein prepared in step (iii) for G-CSF activity and immunogenicity; and (v) selecting a modified G-CSF protein evaluated in step (iv) that has substantially the same therapeutic G-CSF biological activity as, but substantially less immunogenicity than, human G-CSF.