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Details for Patent: 5,811,533

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Details for Patent: 5,811,533

Title: High-affinity oligonucleotide ligands to vascular endothelial growth factor (VEGF)
Abstract:Methods are described for the identification and preparation of nucleic acid ligands to vascular endothelial growth factor (VEGF). Included in the invention are specific RNA ligands to VEGF identified by the SELEX method.
Inventor(s): Gold; Larry (Boulder, CO), Janjic; Nebojsa (Boulder, CO)
Assignee: NeXstar Pharmaceuticals, Inc. (Boulder, CO)
Filing Date:May 19, 1995
Application Number:08/447,169
Claims:1. A nucleic acid ligand to vascular endothelial growth factor (VEGF) identified according to the method comprising:

a) contacting a candidate mixture of nucleic acids with VEGF, wherein nucleic acids 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 nucleic acids from the remainder of the candidate mixture; and

c) amplifying the increased affinity nucleic acids to yield a ligand-enriched mixture of nucleic acids, whereby a nucleic acid ligand to VEGF may be identified.

2. A purified and isolated non-naturally occurring nucleic acid ligand to VEGF.

3. The nucleic acid ligand of claim 2 which is a ribonucleic acid.

4. The RNA ligand of claim 3 wherein said ligand is selected from the group consisting of the sequences set forth in FIG. 2.

5. The RNA ligand of claim 3 wherein said ligand has been chemically modified at the ribose and/or phosphate and/or base positions.

6. The modified RNA ligand of claim 5 wherein said ligand is comprised of 2'-amino (2'-NH.sub.2) modified nucleotides.

7. The modified RNA ligand of claim 6 wherein said ligand is selected from the group consisting of the sequences set forth in FIG. 9 and Table 4.

8. The nucleic acid ligand of claim 2 which is a deoxyribonucleic acid.

9. The DNA ligand of claim 8 wherein said ligand is selected from the group consisting of the sequences set forth in Table 8.

10. The nucleic acid ligand of claim 1 wherein the method further comprises modifying the nucleic acid ligand wherein said modifying comprises adding a moiety that decreases the activity of endonucleases or exonucleases on the nucleic acid ligand relative to the unmodified nucleic acid ligand, without adversely affecting the binding affinity of said nucleic acid ligand.

11. The nucleic acid ligand of claim 10 wherein said moiety comprises a phosphorothioate nucleotide.

12. The nucleic acid ligand of claim 1 wherein the method further comprises modifying the nucleic acid ligand wherein said modifying comprises substituting nucleotide residues, wherein said residues may be chemically modified, and wherein the modification decreases the activity of endonucleases or exonucleases relative to the unmodified nucleic acid ligand, without adversely affecting the binding affinity of said nucleic acid ligand.

13. The nucleic acid of claim 12 wherein said chemically modified residue comprises a 2'-O-Methyl (2'OMe) nucleotide.

14. A modified nucleic acid ligand of VEGF, wherein said ligand is modified to comprise the addition of a moiety to a nucleic acid ligand that decreases the activity of endonucleases or exonucleases on the nucleic acid ligand, without adversely affecting the binding affinity of said nucleic acid ligand.

15. The nucleic acid ligand of claim 14 wherein said moiety comprises a phosphorothioate nucleotide.

16. A modified nucleic acid ligand of VEGF, wherein the modification comprises the substitution of nucleotide residues of the nucleic acid ligand, wherein said residues may be chemically modified, and wherein said modification decreases the activity of endonucleases or exonucleases on the nucleic acid ligand relative to the unmodified nucleic acid ligand, without adversely affecting the binding affinity of said nucleic acid ligand.

17. The nucleic acid ligand of claim 16, wherein said chemically modified residue comprises a 2'-O-Methyl (2'-OMe) nucleotide.

18. The modified RNA ligand of claim 5 wherein said ligand is comprised of a 2'-O-Methyl (2'-OMe) modified nucleotide.
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