Vascular endothelial growth factor variants
AssigneeScios, Inc.
InventorsN. Stephen Pollitt, Judith A. Abraham
Life of this patent
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Abstract
The invention is directed to a method of enhancing the biological activity of vascular endothelial growth factors (VEGF). The invention further concerns certain VEGF variants having enhanced biological activity, methods and means for preparing these variants, and pharmaceutical compositions comprising them. In a further aspect, the invention concerns methods of treatment using, and articles of manufacture containing such VEGF variants.
Claims17
A method of enhancing the biological activity of a vascular endothelial growth factor (VEGF) originally having a cysteine (C) residue at a position corresponding to amino acid position 116 of the 121 amino acids long native mature human VEGF (hVEGF121) (position 142 of SEQ ID NO:
- 2) and an N-linked glycosylation site at a position corresponding to amino acid positions 75-77 of hVEGF121 (positions 101-103 of SEQ ID NO:
- 2), comprising (a) eliminating said cysteine (C) residue and removing said N-linked glycosylation site by site-directed mutagenesis of the encoding nucleic acid sequence to produce a VEGF variant, said variant having at least 85% sequence identity with the native mature hVEGF121 polypeptide (amino acids 27-147 of SEQ ID NO:
- 2), and (b) determining the biological activity of said VEGF variant relative to the said hVEGF121 to confirm the enhancement of biological activity.
A variant of a native vascular endothelial growth factor (VEGF) having a cysteine (C) residue at amino acid position 116 and a glycosylation site at amino acid positions 75-77, comprising the substitution of said cysteine (C) by another amino acid and having said glycosylation site removed, wherein the amino acid numbering follows the numbering of the 121 amino acids long native human VEGF (hVEGF121) (amino acids 27-147 of SEQ ID NO:
- 2), and wherein said variant has at least 85% sequence identity with a mature hVEGF121 polypeptide (amino acids 27-147 of SEQ ID NO:
- 2), and has enhanced biological activity compared to mature hVEGF121 (amino acids 27-147 of SEQ ID NO:
- 2).
An article of manufacture comprising a VEGF variant (a) having a cysteine (C) residue at amino acid position 116 substituted by another amino acid, and a glycosylation site at amino acid positions 75-77 removed by site-directed mutagenesis of the encoding nucleic acid, where the amino acid numbering follows the numbering of the 121 amino acids long native human VEGF (hVEGF121) (amino acids 27-147 of SEQ ID NO:
- 2), (b) having at least 85% sequence identity with the mature hVEGF121 polypeptide (amino acids 27-147 of SEQ ID NO:
- 2), and (c) having enhanced biological activity relative to said mature hVEGF121 (amino acids 27-147 of SEQ ID NO:
- 2);
- a container;
- and
- a label or package insert comprising instructions for administration of said VEGF variant.
Dependencies are inferred from the claim text.