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Details for Patent: 6,001,988

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Details for Patent: 6,001,988

Title: High affinity nucleic acid ligands to lectins
Abstract:This invention discloses high-affinity oligonucleotide ligands to lectins, specifically nucleic acid ligands having the ability to bind to the lectins, wheat germ agglutinin, L-selectin, E-selectin and P-selectin. Also disclosed are the methods for obtaining such ligands.
Inventor(s): Parma; David H. (Boulder, CO), Hicke; Brian J. (Boulder, CO), Bridonneau; Philippe (Boulder, CO), Gold; Larry (Boulder, CO)
Assignee: NeXstar Pharmaceuticals, Inc. (Boulder, CO)
Filing Date:Jun 07, 1995
Application Number:08/472,256
Claims:1. A purified and isolated non-naturally occurring nucleic acid ligand to a lectin selected from the group consisting of wheat germ agglutinin, serum mannose binding protein, and a selectin.

2. The nucleic acid ligand of claim 1 which is a non-naturally occurring nucleic acid ligand having a specific binding affinity for said lectin, such lectin being a three dimensional chemical structure other than a polynucleotide that binds to said nucleic acid ligand through a mechanism which predominantly depends on Watson/Crick base pairing or triple helix binding, wherein said nucleic acid ligand is not a nucleic acid having the known physiological function of being bound by said lectin.

3. The nucleic acid ligand of claim 1 wherein said lectin is wheat germ agglutinin.

4. The nucleic acid ligand to wheat germ agglutinin of claim 3 wherein said nucleic acid ligand is a ribonucleic acid (RNA).

5. The nucleic acid ligand to wheat germ agglutinin of claim 4 wherein said ligand is an RNA ligand selected from the group consisting of the nucleotide sequences set forth in Table II.

6. The nucleic acid ligand of claim 5 wherein said ligand is selected from the group consisting of SEQ ID NOS: 4-55.

7. The nucleic acid ligand of claim 4 wherein said ligand comprises sequences selected from the group consisting of SEQ ID NOS: 56-63.

8. The nucleic acid ligand to wheat germ agglutinin of claim 3 wherein said ligand is substantially homologous to and has substantially the same ability to bind said wheat germ agglutinin as a ligand selected from the group consisting of the sequences set forth in Table II.

9. The nucleic acid ligand to wheat germ agglutinin of claim 3 wherein said ligand has substantially the same structure and has substantially the same ability to bind said wheat germ agglutinin as a ligand selected from the group consisting of the sequences set forth in Table II.

10. The nucleic acid ligand of claim 1 wherein said lectin is a selectin.

11. The nucleic acid ligand of claim 10 wherein said selectin is selected from the group consisting of L-selectin, E-selectin and P-selectin.

12. The nucleic acid ligand of claim 11 wherein said selectin is L-selectin.

13. The nucleic acid ligand to L-selectin of claim 12 wherein said nucleic acid ligand is a ribonucleic acid (RNA).

14. The nucleic acid ligand to L-selectin of claim 12 wherein said nucleic acid ligand is a deoxyribonucleic acid (DNA).

15. The nucleic acid ligand to L-selectin of claim 13 wherein said ligand is an RNA ligand selected from the group consisting of the nucleotide sequences set forth in Table VIII.

16. The nucleic acid ligand of claim 15 wherein said ligand is selected from the group consisting of SEQ ID NOS: 67-117.

17. The nucleic acid ligand of claim 13 wherein said ligand comprises sequences selected from the group consisting of SEQ ID NOS: 118-125.

18. The nucleic acid ligand to L-selectin of claim 14 wherein said ligand is a DNA ligand selected from the group consisting of the nucleotide sequences set forth in Table XII.

19. The nucleic acid ligand of claim 18 wherein said ligand is selected from the group consisting of SEQ ID NOS: 129-167.

20. The nucleic acid ligand of claim 14 wherein said ligand comprises sequences selected from the group consisting of SEQ ID NOS: 168-171.

21. The nucleic acid ligand of claim 1 wherein said ligand is a ribonucleic acid ligand.

22. The nucleic acid ligand of claim 1 wherein said ligand is a deoxyribonucleic acid.

23. The nucleic acid ligand of claim 1 wherein said ligand comprises 2'-amino (2'-NH.sub.2) modified ligands, 2'-fluoro (2'-F) modified ligands and 2'-O-methyl (2'-OMe) modified ligands.

24. A nucleic acid ligand to a lectin selected from the group consisting of wheat germ agglutinin, serum mannose binding protein, L-selectin, E-selectin and P-selectin, identified according to the method comprising:

a) contacting a candidate mixture of nucleic acids with said lectin, wherein nucleic acids having an increased affinity to said lectin 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 mixture of nucleic acids enriched for nucleic acid sequences with increased affinity and specificity for binding to said lectin, whereby nucleic acid ligands of said lectin may be identified.

25. The nucleic acid ligand to L-selectin of claim 12 wherein said ligand is substantially homologous to and has substantially the same ability to bind said L-selectin as a ligand selected from the group consisting of the sequences set forth in Table VIII.

26. The nucleic acid ligand to L-selectin of claim 12 wherein said ligand has substantially the same structure and substantially the same ability to bind said L-selectin as a ligand selected from the group consisting of the sequences set forth in Table VIII.

27. The nucleic acid ligand to L-selectin of claim 12 wherein said ligand is substantially homologous to and has substantially the same ability to bind said L-selectin as a ligand selected from the group consisting of the sequences set forth in Table XII.

28. The nucleic acid ligand to L-selectin of claim 12 wherein said ligand has substantially the same structure and substantially the same ability to bind said L-selectin as a ligand selected from the group consisting of the sequences set forth in Table XII.
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