Nucleic acid ligands of tissue target

• US 6,127,119 A
• Filed: 11/21/1997
• Issued: 10/03/2000
• Est. Priority Date: 06/11/1990
• Status: Expired due to Fees

• Current Assignee: Gilead Sciences Inc., Schering Ag (Merck & Co., Inc.)
• Original Assignee: NeXstar Pharmaceuticals, Inc. (Gilead Sciences Inc.)
• Inventors: Gold, Larry, Speck, Ulrich, Stephens, Andrew
• Primary Examiner(s): ZITOMER, STEPHANIE W
• Application Number:
  US08/976,413
• Time in Patent Office: 1,047 Days
• Field of Search: 435/6, 435/91.2, 536/24.3, 536/24.31, 536/25.4, 514/44, 424/1.73, 935/77, 935/78
• US Class Current: 435/6.11
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• 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: C07K 14/005 from viruses
• CPC Class Codes: C07K 14/72 for hormones for neuromediators C07K14/70571
• CPC Class Codes: C07K 7/22 Tachykinins, e.g. Eledoisins , Substance P; Related peptides
• 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 2310/16 Aptamers
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/344 Position-specific modifications, e.g. on every purine, at the 3'-end
• CPC Class Codes: C12N 2310/53 partially self-complementary or closed
• CPC Class Codes: C12N 2740/16222 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• 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: C40B 40/00 : Libraries per se, e.g. arrays, mixtures
• CPC Class Codes: F02B 2075/027 : four
• CPC Class Codes: G01N 2333/16 : HIV-1, HIV-2
• 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 2333/503 : basic FGF [bFGF]
• 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 2333/96436 : Granzymes
• CPC Class Codes: G01N 2333/96455 : Kallikrein (3.4.21.34; 3.4.21.35)
• CPC Class Codes: G01N 2333/96486 : Metalloendopeptidases (3.4.24)
• 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 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.

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'"'"'-gggagacaagaauaaacgcucaaUCAAUCUCGGACUAGACUAACGACCUUGGUUGACGCUCAuucgacaggaggcucacaacagga-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.