G-CSF conjugates
AssigneeMaxygen Holdings Ltd.
InventorsTorben Lauesgaard Nissen, Kim Vilbour Andersen, Christian Karsten Hansen, Jan Moller Mikkelsen, Hans Thalsgaard Schambye
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Abstract
Polypeptide conjugates with G-CSF activity comprising a polypeptide having at least one introduced lysine residue and at least one removed lysine residue compared to the sequence of human G-CSF, and which are conjugated to 2-6 polyethylene glycol moieties. The conjugates have a low in vitro bioactivity, a long in vivo half-life, a reduced receptor-mediated clearance, and provide a more rapid stimulation of production of white blood cells and neutrophils than non-conjugated recombinant human G-CSF.
Claims20
A polypeptide conjugate exhibiting G-CSF activity, comprising a polypeptide comprising at least one substitution selected from the group consisting of K16R/Q, K34R/Q and K40R/Q, and at least one substitution selected from the group consisting of T105K and S159K, relative to the amino acid sequence of hG-CSF shown in SEQ ID NO:
- 1 or in a corresponding position relative to an amino acid sequence having at least 80% sequence identity with SEQ ID NO:
- 1, the conjugate having 2-6 polyethylene glycol moieties with a molecular weight of about 1000-10,000 Da attached to attachment groups of the polypeptide.
A method for preparing a G-CSF conjugate which provides a reduced duration of neutropenia compared to hG-CSF, the method comprising preparing a polypeptide comprising an amino acid sequence that differs in at least one amino acid residue from the amino acid sequence of hG-CSF shown in SEQ ID NO:
- 1, and attaching a non-polypeptide moiety to each of at least two attachment groups of said polypeptide to result in a conjugate having an in vitro bioactivity in the range of about 2-30% of the bioactivity of non-conjugated hG-CSF as determined by the luciferase assay described herein.
Dependencies are inferred from the claim text.