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Claims for Patent: 6,051,698

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Claims for Patent: 6,051,698

Title: Vascular endothelial growth factor (VEGF) nucleic acid ligand complexes
Abstract:This invention discloses a method for preparing a complex comprised of a VEGF Nucleic Acid Ligand and a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound by identifying a VEGF Nucleic Acid Ligand by SELEX methodology and associating the VEGF Nucleic Acid Ligand with a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound. The invention further discloses Complexes comprising one or more VEGF Nucleic Acid Ligands in association with a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound. The invention further includes a Lipid construct comprising a VEGF Nucleic Acid Ligand or Complex and methods for making the same.
Inventor(s): Janjic; Nebojsa (Boulder, CO), Gold; Larry (Boulder, CO), Schmidt; Paul (Niwot, CO), Vargeese; Chandra (Thornton, CO)
Assignee:
Application Number:08/897,351
Patent Claims: 1. A purified and isolated non-naturally occurring RNA ligand to Vascular Endothelial Growth Factor wherein said ligand is comprised of 2'fluoro (2'F)-modified nucleotides.

2. A method for improving the pharmacokinetic properties of VEGF Nucleic Acid Ligand in an occular application comprising covalently linking a Non-Immunogenic, High Molecular Weight compound to a VEGF Nucleic Acid Ligand to form a complex comprised of a VEGF Nucleic Acid Ligand and a Non-Immunogenic, High Molecular Weight Compound.

3. The RNA ligand to VEGF of claim 1 wherein said ligand is selected from the group consisting of the sequences set forth in Tables 1-4 (SEQ ID NOS: 10-86).

4. The RNA ligand of claim 3 wherein said ligand is substantially homologous to and has substantially the same ability to bind VEGF as a ligand selected from the group consisting of the sequences set forth in Tables 1-4 (SEQ ID NOS: 10-86).

5. The RNA ligand of claim 3 wherein said ligand is substantially the same structure and substantially the same ability to bind VEGF as a ligand selected from the group consisting of the sequences set forth in Tables 1-4 (SEQ ID NOS: 10-86).

6. The RNA ligand to VEGF of claim 1 identified according to the method comprising:

a) contacting a Candidate Mixture of RNA with VEGF, wherein the RNA having an increased affinity to VEGF relative to the Candidate Mixture may be partitioned from the remainder of the Candidate Mixture;

b) partitioning the increased affinity RNA from the remainder of the Candidate Mixture; and

c) amplifying the increased affinity RNA to yield a mixture of RNA enriched for RNA having an increased affinity for VEGF, whereby RNA Ligands of VEGF are identified.

7. The RNA ligand to VEGF of claim 3 identified according to the method comprising:

a) contacting a Candidate Mixture of RNA with VEGF, wherein the RNA having an increased affinity to VEGF relative to the Candidate Mixture may be partitioned from the remainder of the Candidate Mixture;

b) partitioning the increased affinity RNA from the remainder of the Candidate Mixture; and

c) amplifying the increased affinity RNA to yield a mixture of RNA enriched for RNA having an increased affinity for VEGF; whereby RNA Ligands of VEGF are identified.

8. A Complex comprised of the RNA ligand to VEGF of claim 1 and a Non-Immunogenic, High Molecular Weight Compound.

9. The Complex of claim 8 further comprising a Linker between said ligand and said Non-Immunogenic, High Molecular Weight Compound.

10. The Complex of claim 8 wherein said Non-Immunogenic, High Molecular Weight Compound is a Polyalkylene Glycol.

11. The Complex of claim 10 wherein said Polyalkylene Glycol is polyethylene glycol.

12. The Complex of claim 11 wherein said polyethylene glycol has a molecular weight of about 10-80 Kd.

13. The Complex of claim 11 wherein said polyethylene glycol has a molecular weight of about 20-45 Kd.

14. The Complex of claim 11 wherein said Complex is ##STR9##

15. The Complex of claim 11 wherein said Complex is

16. The method of claim 2 wherein said Non-Immunogenic, High Molecular Weight Compound is a Polyalkylene Glycol.

17. The method of claim 16 wherein said Polyalkylene Glycol is polyethylene glycol.

18. The method of claim 17 wherein said polyethylene glycol has a molecular weight of about 10-80 Kd.

19. The method of claim 17 wherein said polyethylene glycol has a molecular weight of about 20-45 Kd.

20. The method of claim 19 wherein said complex has the structure

21. The complex of claim 19 wherein said complex is
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