Remodeling and Glycopegylation of Fibroblast Growth Factor (Fgf)
OwnerShawn DeFreesinventor
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Abstract
The present invention relates to mutants of Fibroblast Growth Factor (FGF), particularly FGF-20 and FGF-21, which contain newly introduced N-linked or O-linked glycosylation site(s). The polynucleotide coding sequences for the mutants, expression cassettes comprising the coding sequences, cells expressing the mutants, and methods for producing the mutants are also disclosed. Further disclosed are pharmaceutical compositions comprising the mutants and method for using the mutants.
Claims51
A Fibroblast Growth Factor (FGF) conjugate comprising:
- a FGF peptide;
- and
- a modifying group, wherein said modifying group is covalently attached to said peptide at a preselected glycosyl or amino acid residue of said peptide via an intact glycosyl linking group.
An isolated nucleic acid comprising a polynucleotide sequence encoding a mutant Fibroblast Growth Factor, wherein the mutant Fibroblast Growth Factor comprises a newly introduced N-linked or O-linked glycosylation site that does not exist in the corresponding wild-type Fibroblast Growth Factor.
A mutant Fibroblast Growth Factor, comprising a newly introduced N-linked or O-linked glycosylation site that does not exist in the corresponding wild-type Fibroblast Growth Factor.
A method for making a mutant Fibroblast Growth Factor, which comprises a newly introduced N-linked or O-linked glycosylation that does not exist in the corresponding wild-type Fibroblast Growth Factor, comprising the steps of:
- (a) recombinantly producing the mutant Fibroblast Growth Factor;
- and
- (b) glycosylating the mutant Fibroblast Growth Factor at the newly introduced glycosylation site
- wherein
- said glycosylating is a cell free, in vitro process.
A pharmaceutical composition comprising an effective amount of a mutant Fibroblast Growth Factor, which comprises a newly introduced N-linked or O-linked glycosylation that does not exist in the corresponding wild-type Fibroblast Growth Factor.
A method for treating Fibroblast Growth Factor deficiency in a patient, comprising the step of administering an effective amount of a mutant Fibroblast Growth Factor to the patient, wherein the mutant Fibroblast Growth Factor comprises a newly introduced N-linked or O-linked glycosylation that does not exist in the corresponding wild-type Fibroblast Growth Factor.
A method for making a glycoconjugate of a mutant Fibroblast Growth Factor, which comprises a newly introduced N-linked or O-linked glycosylation that does not exist in the corresponding wild-type Fibroblast Growth Factor, comprising the steps of:
- (a) recombinantly producing the mutant Fibroblast Growth Factor, and
- (b) enzymatically glycosylating the mutant Fibroblast Growth Factor with a modified sugar at the newly introduced glycosylation site
- wherein
- said glycosylating is a cell free, in vitro process.
Dependencies are inferred from the claim text.