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|Title:||High-affinity oligonucleotide ligands to vascular endothelial growth factor (VEGF)|
|Abstract:||Methods are described for the identification and preparation of nucleic acid ligand 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:||Gilead Sciences, Inc. (Foster City, CA)|
|Filing Date:||May 18, 2001|
|Claims:||1. A method of inhibiting binding of vascular endothelial growth factor (VEGF) to its receptor, comprising: a) providing a nucleic acid ligand to VEGF; and b) contacting the nucleic acid ligand with said VEGF under conditions where a VEGF:nucleic acid binding pair is formed, whereby binding of VEGF to its receptor is inhibited. |
2. The method of claim 1, wherein the nucleic acid ligand to VEGF is identified according to a method comprising the steps of: a) preparing a candidate mixture of nucleic acids; b) contacting the candidate mixture 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; c) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and d) amplifying the increased affinity nucleic acids to yield a ligand-enriched mixture of nucleic acids, whereby a VEGF nucleic acid ligand may be identified.
3. The method of claim 1, wherein said VEGF nucleic acid ligand is a ribonucleic acid.
4. The method of claim 1, wherein said VEGF nucleic acid ligand is a deoxyribonucleic acid.
5. The method of claim 1, wherein said VEGF nucleic acid ligand comprises modified nucleotides.
6. The method of claim 5, wherein said VEGF ligand is comprised of 2'-amino (2'-NH.sub.2) modified nucleotides and 2'-O-methyl (2'-OMe) modified nucleotides.
7. The method of claim 1, wherein said VEGF nucleic acid ligand is modified with 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.
8. The method of claim 7, wherein said moiety comprises a phosphorothioate nucleotide.
9. The method of claim 1, wherein said VEGF nucleic acid ligand is modified by a method comprising 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.
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