|Title:||High-affinity RNA ligands for basic fibroblast growth factor|
|Abstract:||Methods are described for the identification and preparation of nucleic acid ligand solutions to basic fibroblast growth factor (bFGF). Included in the invention are nucleic acid ligands to bFGF which are inhibitors of bFGF and 2'-amino-modified RNA ligands to bFGF.|
|Inventor(s):||Janjic; Nebojsa (Boulder, CO), Gold; Larry (Boulder, CO)|
|Assignee:||NeXstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Feb 06, 1995|
|Claims:||1. A nucleic acid ligand to basic fibroblast growth factor (bFGF) identified according to the method comprising: |
a) preparing a candidate mixture of nucleic acids;
b) contacting the candidate mixture with bFGF, wherein nucleic acids having an increased affinity to bFGF relative to the candidate mixture may be partitioned from the remainder of the candidate mixture;
c) partitioning the increased affinity nucleic acid ligands from the remainder of the candidate mixture; and
d) amplifying the increased affinity nucleic acid ligands for bFGF to yield a mixture of nucleic acids enriched for sequences with a higher affinity to bFGF, whereby a nucleic acid ligand of bFGF may be identified.
2. The nucleic acid ligand of claim 1 comprising a single stranded nucleic acid.
3. The nucleic acid ligand of claim 1 comprised of RNA.
4. The nucleic acid ligand of claim 3 comprised of RNA wherein all pyrimidines are 2'-deoxy-2'-NH.sub.2 pyrimidines.
5. A purified and isolated non-naturally occurring nucleic acid ligand to bFGF.
6. The nucleic acid ligand of claim 5 wherein said ligand is a RNA ligand.
7. The RNA ligand of claim 6 wherein said RNA ligand is selected from the group consisting of the nucleotide sequences set forth in FIGS. 4 and 5 and Tables IV and VIII, (SEQ ID NO:48-69 and SEQ ID NO:101-185, respectively).
8. The RNA ligand of claim 6 wherein said RNA ligand is an inhibitor of bFGF.
9. The nucleic acid ligand of claim 1 wherein the method of identifying further comprises
e) repeating steps b), c) and d).
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