|Title:|| Ligands of thrombin|
|Abstract:||Methods are described for the identification of nuclei acid ligand solutions to thrombin. The present invention utilizes the SELEX (Systematic Evolution of Ligands for EXponential enrichment) method for identifying and preparing RNA ligands to thrombin. Further included in the present invention are modified nucleotide sequences based on the sequences of the RNA ligands identified. The modified RNA ligands to thrombin exhibit increased in vivo stability.|
|Inventor(s):|| Gold; Larry (Boulder, CO), Tasset; Diane (Boulder, CO) |
|Assignee:|| Nexstar Pharmaceuticals, Inc. (Boulder, CO) |
|Filing Date:||Nov 06, 1992|
|Claims:||1. A method for identifying nucleic acid ligands to thrombin comprising: |
a) preparing a candidate mixture of RNA nucleic acids;
b) contacting the candidate mixture with thrombin, wherein RNA nucleic acids having an increased affinity to thrombin relative to the candidate mixture RNA nucleic acids may be partitioned from the remainder of the candidate mixture;
c) partitioning the increased affinity RNA nucleic acids from the remainder of the candidate mixture; and
d) amplifying the increased affinity nucleic acids, in vitro, to yield a ligand-enriched mixture of RNA nucleic acids, whereby RNA nucleic acid ligands to thrombin may be identified.
2. The method of claim 1 further comprising
e) repeating steps b), c) and d).
3. The method of claim 1 wherein said candidate mixture of RNA nucleic acids is comprised of single stranded nucleic acids.
4. A nucleic acid ligand to thrombin identified according to the method of claim 1.
5. The nucleic acid ligand of claim 4 being a single stranded nucleic acid.
6. A purified and isolated non-naturally occurring RNA ligand to thrombin wherein the nucleotide sequence of said RNA ligand is selected from the group consisting of the nucleotide sequences set forth in FIG. 1.
7. The RNA ligand of claim 6 consisting of the nucleotide sequence (SEQ ID NO:2):
8. The RNA ligand of claim 6 consisting of having the nucleotide sequence (SEQ ID NO:3):
9. The RNA ligand of claim 6 identified by the method of claim 1.