High affinity nucleic acid ligands to lectins

• US 7,399,752 B2
• Filed: 04/07/2003
• Issued: 07/15/2008
• Est. Priority Date: 06/11/1990
• Status: Expired due to Fees

• Current Assignee: Gilead Sciences Inc.
• Original Assignee: Gilead Sciences Inc.
• Inventors: Hicke, Brian, Bridonneau, Philippe, Gold, Larry, Parma, David H.
• Primary Examiner(s): Kim; Young J.
• Assistant Examiner(s): WOOLWINE, SAMUEL C
• Application Number:
  US10/409,627
• Publication Number:
  US 20040043923A1
• Time in Patent Office: 1,926 Days
• Field of Search: None
• US Class Current: 514/44.00R
• CPC Class Codes: A61K 47/547 Chelates, e.g. Gd-DOTA or Zinc-amino acid chelates; Chelate-forming compounds, e.g. DOTA or ethylenediamine being covalently linked or complexed to the pharmacologically- or therapeutically-active agent
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07K 14/001 by chemical synthesis
• CPC Class Codes: C12N 15/1048 SELEX
• CPC Class Codes: C12N 15/115 Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
• CPC Class Codes: C12N 2310/13 Decoys
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/53 partially self-complementary or closed
• CPC Class Codes: C12N 9/1276 RNA-directed DNA polymerase (2.7.7.49), i.e. reverse transcriptase or telomerase
• CPC Class Codes: C12Q 1/37 involving peptidase or proteinase
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: C12Q 1/6811 Selection methods for production or design of target specific oligonucleotides or binding molecules
• CPC Class Codes: C12Q 1/70 involving virus or bacteriophage immunoassay for viruses G01N33/56983
• CPC Class Codes: C12Q 2525/101 incorporating non-naturally occurring nucleotides, e.g. inosine
• CPC Class Codes: C40B 40/00 Libraries per se, e.g. arrays, mixtures
• CPC Class Codes: F02B 2075/027 four
• CPC Class Codes: G01N 2333/163 : Regulatory proteins, e.g. tat, nef, rev, vif, vpu, vpr, vpt, vpx
• CPC Class Codes: G01N 2333/4724 : Lectins
• CPC Class Codes: G01N 2333/575 : Hormones derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin G01N2333/665, corticotropin G01N2333/695
• CPC Class Codes: G01N 2333/62 : Insulins
• CPC Class Codes: G01N 2333/7056 : Selectin superfamily, e.g. LAM-1, GlyCAM, ELAM-1, PADGEM
• CPC Class Codes: G01N 2333/8125 : Alpha-1-antitrypsin
• CPC Class Codes: G01N 2333/96433 : Serine endopeptidases (3.4.21)
• CPC Class Codes: G01N 2333/96455 : Kallikrein (3.4.21.34; 3.4.21.35)
• CPC Class Codes: G01N 2333/966 : Elastase
• CPC Class Codes: G01N 2333/9726 : Tissue plasminogen activator
• CPC Class Codes: G01N 2333/974 : Thrombin
• CPC Class Codes: G01N 2333/976 : Trypsin; Chymotrypsin
• CPC Class Codes: G01N 33/53 : Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/531 : Production of immunochemical test materials
• CPC Class Codes: G01N 33/532 : Production of labelled immunochemicals
• CPC Class Codes: G01N 33/535 : with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
• CPC Class Codes: G01N 33/56988 : HIV or HTLV
• CPC Class Codes: G01N 33/68 : involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 33/76 : Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors

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.

Claims

1. A method of inhibiting binding between L-selectin and a carbohydrate ligand of said L-selectin, comprising the step of contacting a nucleic acid ligand to said L-selectin with said L-selectin wherein said nucleic acid ligand to L-selectin is selected from the group consisting of SEQ ID NO:

67-117, 129-196, and 293-388.

2. A method of inhibiting binding between P-selectin and a carbohydrate ligand of said P-selectin, comprising the step of contacting a nucleic acid ligand to said P-selectin with said P-selectin wherein said nucleic acid ligand to P-selectin is selected from the group consisting of SEQ ID NO:

199-247, and 251-290.

3. A method of modulating leukocyte adhesion or migration, comprising inhibiting binding between a P- or L-selectin on a first cell and a ligand to said selectin on a second cell, said inhibiting comprising the step of permitting a nucleic acid ligand to the selectin to interact with the first cell, wherein the first cell is a leukocyte selected from the group consisting of neutrophils and monocytes, the second cell is a vascular endothelial cell and wherein said nucleic acid ligand is selected from the group consisting of SEQ ID NO:

67-117, 129-196, 199-247, 251-290 and 293-388.

4. A method for modulating platelet adhesion, comprising inhibiting binding between a P-selectin on a first cell and a ligand to said P-selectin on a second cell, said inhibiting comprising the step of permitting a nucleic acid ligand to said P-selectin to interact with the first cell, wherein the first cell is a platelet, wherein said nucleic acid ligand to said P-selectin is selected from the group consisting of SEQ ID NO:

199-247 and 251-290.

5. A method of modulating leukocyte adhesion or migration, comprising inhibiting binding between an L-selectin on a first cell and a ligand to said L-selectin on a second cell, said inhibiting comprising the step of permitting a nucleic acid ligand to said L-selectin to interact with the first cell, wherein the first cell is a leukocyte selected from the group consisting of lymphocytes, granulocytes, neutrophils and monocytes, the second cell is a vascular endothelial cell or a lymphatic cell and wherein said nucleic acid ligand to said L-selectin is selected from the group consisting of SEQ ID NO:

67-117, 129-196 and 293-388.

6. A method of modulating lymphocyte adhesion, migration or activation comprising inhibiting binding between an L-selectin on a first cell and a ligand to said L-selectin on a second cell, said inhibiting comprising the step of permitting a nucleic acid ligand to said L-selectin to interact with the first cell, wherein the first cell is a lymphocyte, the second cell is a vascular endothelial cell and wherein said nucleic acid ligand is selected from the group consisting of SEQ ID NO:

67-117, 129-196 and 293-388.

7. A method of inhibiting binding between P-selectin and a carbohydrate ligand of said P-selectin, comprising the step of contacting a nucleic acid ligand to said P-selectin with said P-selectin wherein said nucleic acid ligand to P-selectin comprises SEQ ID NO:

223.

8. The method of claim 7 wherein SEQ ID NO:

223 further comprises at least one 2′

-O-methyl purine.

9. The method of claim 7 wherein SEQ ID NO:

223 further comprises 2′

-OMe purines at positions 7, 8, 9, 15, 27, 28 and 31.

10. The method of claim 9 wherein SEQ ID NO:

223 further comprises a 2′

-OMe purine at one or more of the following positions 13, 14, 16, 18, 21, 22, 24 and 30.

11. The method of claim 9 wherein SEQ ID NO:

223 further comprises 2′

-OMe purines at positions 13, 14, 18, 21, 22, and 24.

12. A method of modulating leukocyte adhesion or migration, comprising inhibiting binding between a P-selectin on a first cell and a ligand to said P-selectin on a second cell, said inhibiting comprising the step of permitting a nucleic acid ligand to the selectin to interact with the first cell, wherein the first cell is a leukocyte selected from the group consisting of neutrophils and monocytes, the second cell is a vascular endothelial cell and wherein said nucleic acid ligand comprises SEQ ID NO:

223.

13. The method of claim 12 wherein SEQ ID NO:

223 further comprises at least one 2′

-O-methyl purine.

14. The method of claim 12 wherein SEQ ID NO:

223 further comprises 2′

-OMe purines at positions 7, 8, 9, 15, 27, 28 and 31.

15. The method of claim 14 wherein SEQ ID NO:

223 further comprises a 2′

-OMe purine at one or more of the following positions 13, 14, 16, 18, 21, 22, 24 and 30.

16. The method of claim 14 wherein SEQ ID NO:

223 further comprises 2′

-OMe purines at positions 13, 14, 18, 21, 22, and 24.

17. A method for modulating platelet adhesion, comprising inhibiting binding between a P-selectin on a first cell and a ligand to said P-selectin on a second cell, said inhibiting comprising the step of permitting a nucleic acid ligand to said P-selectin to interact with the first cell, wherein the first cell is a platelet, wherein said nucleic acid ligand to said P-selectin comprises SEQ ID NO:

223.

18. The method of claim 17 wherein SEQ ID NO:

223 further comprises at least one 2′

-O-methyl purine.

19. The method of claim 17 wherein SEQ ID NO:

223 further comprises 2′

-OMe purines at positions 7, 8, 9, 15, 27, 28 and 31.

20. The method of claim 19 wherein SEQ ID NO:

223 further comprises a 2′

-OMe purine at one or more of the following positions 13, 14, 16, 18, 21, 22, 24 and 30.

21. The method of claim 19 wherein SEQ ID NO:

223 further comprises 2′

-OMe purines at positions 13, 14, 18, 21, 22, and 24.