Claims
- 1. A method of inducing angiogenesis comprising:
administering an effective amount of a VEGF variant to a patient in need thereof, wherein the VEGF variant is encoded by a nucleic acid sequence encoding a vascular endothelial growth fact (VEGF) in which 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) has been removed 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) has been removed, said variant having at least 85% sequence identity with the native mature hVEGF121 polypeptide (amino acids 27-147 of SEQ ID NO:2).
- 2. The method of claim 1, wherein angiogenesis is induced to prevent or repair injury to blood vessels.
- 3. The method of claim 2, wherein said injury is associated with microvascular angiopathy.
- 4. The method of claim 3, wherein said microvascular angiopathy is thrombotic microangiopathy (TMA).
- 5. The method of claim 2, wherein said injury is associated with hemolytic uremic syndrom (HUS).
- 6. The method of claim 1, wherein the cysteine (C) residue is substituted by a different amino acid residue.
- 7. The method of claim 6, wherein the different amino acid residue is a serine (S) residue.
- 8. The method of claim 6, wherein, apart from the substitution at position 116 and the removal of the glycosylation site at position 75-77, the VEGF variant retains the amino acid sequence of mature hVEGF121 (amino acids 27-147 of SEQ ID NO:2)
- 9. The method of claim 1, wherein removing the N-linked glycosylation site from the encoding nucleic acid sequence to produce a VEGF variant comprises removing the N-linked glycosylation site using site-directed mutagenesis.
- 10. The method of claim 9, wherein removing the N-linked glycosylation site from the encoding nucleic acid sequence to produce a VEGF variant comprises substituting at least one residue in the asparagine-isoleucine-threonine (N-I-T) glycosylation site at position 75-77.
- 11. The method of claim 10, wherein asparagine (N) at position 75 is substituted by another amino acid.
- 12. The method of claim 10, wherein the another amino acid is glutamine (Q).
- 13. The method of claim 1, wherein the VEGF variant is C116S hVEGF121.
- 14. The method of claim 1, wherein the VEGF variant additionally comprises an N-terminal substitution compared to a native VEGF (hVEGF).
- 15. The method of claim 14, wherein said substitution is within the first 10 amino acids of a hVEGF.
- 16. The method of claim 15, wherein said substitution is at amino acid position 5 of a hVEGF.
- 17. The method of claim 16, wherein said substitution is X5D, wherein X is an amino acid at position 5 of the hVEGF.
RELATED APPLICATIONS
[0001] This application is a divisional of application Ser. No. 09/574,708, filed May 18, 2000, now U.S. Pat. No.______ , which claims the benefit of priority to Application Ser. No. 60/135,312, filed May 20, 1999, both of which are hereby incorporated by reference in their entirety.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60135312 |
May 1999 |
US |
Divisions (1)
|
Number |
Date |
Country |
Parent |
09574708 |
May 2000 |
US |
Child |
10268447 |
Oct 2002 |
US |