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Details for Patent: 6,127,119

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Details for Patent: 6,127,119

Title: Nucleic acid ligands of tissue target
Abstract:This invention discloses high-affinity oligonucleotide ligands to complex tissue targets, specifically nucleic acid ligands having the ability to bind to complex tissue targets, and the methods for obtaining such ligands. Tissue targets comprise cells, subcellular components, aggregates or cells, collections of cells, and higher ordered structures. Specifically, nucleic acid ligands to blood vessels are described.
Inventor(s): Stephens; Andrew (Boulder, CO), Gold; Larry (Boulder, CO), Speck; Ulrich (Berlin, DE)
Assignee: NeXstar Pharmaceuticals, Inc. (Boulder, CO)
Filing Date:Nov 21, 1997
Application Number:08/976,413
Claims:1. A method for identifying a nucleic acid ligand to a blood vessel comprising:

a) preparing a candidate mixture of nucleic acid sequences;

b) contacting said candidate mixture of nucleic acids with said blood vessel, wherein nucleic acids having an increased affinity to the blood vessel 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 mixture of nucleic acids enriched for nucleic acid sequences with relatively higher affinity and specificity for binding to said blood vessel, whereby a nucleic acid ligand of said blood vessel 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 is comprised of single-stranded nucleic acids.

4. The method of claim 3 wherein said single-stranded nucleic acids are ribonucleic acids.

5. The method of claim 3 wherein said single-stranded nucleic acids are deoxyribonucleic acids.

6. The method of claim 1 wherein said blood vessel is selected from the group consisting of arteries, veins and capillaries.

7. A nucleic acid ligand to a blood vessel identified according to the method of claim 1.

8. The nucleic acid ligand of claim 7 wherein said candidate mixture is comprised of single-stranded nucleic acids.

9. The nucleic acid ligand of claim 7 wherein said single-stranded nucleic acids are deoxyribonucleic acids.

10. The nucleic acid ligand of claim 7 wherein said single-stranded nucleic acids arc ribonucleic acids.

11. The nucleic acid ligand of claim 7 wherein said ligand is a RNA ligand selected from the group consisting of the nucleotide sequences set forth in Table 10 (SEQ ID NOS: 242-258).

12. The nucleic acid ligand of claim 7 wherein said blood vessel is selected from the group consisting of arteries, veins and capillaries.

13. The nucleic acid ligand of claim 12 wherein said blood vessel is an artery.

14. The nucleic acid ligand of claim 13 wherein said ligand is a RNA ligand to an atherosclerotic lesion or plaque in said artery selected from the group consisting of the nucleotide sequences SEQ ID NOS: 260-440.

15. The nucleic acid ligand of claim 7 wherein said ligand further comprises a label.

16. The nucleic acid ligand of claim 7 wherein said ligand comprises a complexing agent.

17. The nucleic acid ligand of claim 16 wherein said complexing agent comprises a label.

18. The nucleic acid ligand of claim 17 wherein label is 99'm-Tc.

19. The nucleic acid ligand of claim 18 having the structure: ##STR1## Wherein the ligand has the nucleotide sequence 5'-gggagacaagaauaaacgcucaaUCAAUCUCGGACUAGACUAACGACCUUGGUUGACGCUCAuucgacagg aggcucacaacagga-3' (SEQ ID NO:354) (SEQ ID NO:355).

20. A purified and isolated non-naturally occurring nucleic acid ligand to a blood vessel.

21. The purified and isolated non-naturally occurring nucleic acid ligand of claim 20 wherein said blood vessel is selected from the group consisting of arteries, veins and capillaries.
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