Tenascin-C nucleic acid ligands

• US 7,005,260 B1
• Filed: 01/28/2000
• Issued: 02/28/2006
• Est. Priority Date: 01/28/2000
• Status: Expired due to Fees

• Current Assignee: Gilead Sciences Inc.
• Original Assignee: Gilead Sciences Inc.
• Inventors: Gold, Larry, Hicke, Brian, Warren, Stephen, Parma, David
• Primary Examiner(s): FORMAN, BETTY J
• Application Number:
  US10/031,193
• Time in Patent Office: 2,223 Days
• Field of Search: 435/6, 435/91.2, 435/7.1, 435/91.1, 536/23.1, 536/24.3, 536/26.6, 536/25.3
• US Class Current: 435/5
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

Methods are described for the identification and preparation of nucleic acid ligands to tenascin-C. Included in the invention are specific RNA ligands to tenascin-C identified by the SELEX method. Further included in the invention are methods for detecting the presence of a disease condition in a biological tissue in which tenascin-C is expressed.

Claims

1. A method for detecting the presence of a disease in a biological tissue which may contain said disease, wherein said disease is characterized by the expression of tenascin-C in said tissue and wherein said disease is selected from the group consisting of cancer, psoriasis, and atherosclerosis, the method comprising:

a) attaching a marker that can be used in in vivo diagnostics to a tenascin-C nucleic acid ligand to form a marker-nucleic acid ligand complex wherein said tenascin-C nucleic acid ligand is selected from the group consisting of SEQ ID NO;

4–

65;

b) exposing said biological tissue which may contain said disease to said marker-nucleic acid ligand complex; and

c) detecting the presence of said disease in said tissue by detecting the presence of said marker-nucleic acid ligand in said tissue.

2. The method of 1 wherein said marker is selected from from the group consisting of radionuclides, fluorophores, magnetic compounds, and biotin.

3. The method of 2 wherein said radionuclide is selected from the group consisting of technetium-99m (Tc-99m), Re-188, Cu-64, Cu-67, F-18, ¹²⁵I, ¹³¹I, ¹¹¹In, ³²P, and ¹⁸⁶Re.

4. The method of 3 wherein said marker is technetium-99m.

5. The method of 4 wherein said tenascin-C nucleic acid ligand comprises a linker.

6. The method of 5 wherein said linker is (CH₂CH₂O)₆.

7. The method of 5, wherein said linker has the structure

8. The method of 1 wherein said tenascin-C nucleic acid ligand is

5′

-B-G667667CG-(CH₂CH₂O)₆-CGUCGCCGU77U667U6UUUU6CUCCCU65

wherein;

all pyrimidines are 2′

F;

6=2′

OMe G;

7=2′

OMe A;

5=3′

—

3′

dT; and

B=linker.

9. The method of 8 wherein said technetium-99m is associated with a chelator.

10. The method of 9, wherein said complex is

11. The method of 10 wherein said complex is

12. The method of 1 further comprising attaching a therapeutic or diagnostic agent to said complex.

13. The method of 1 wherein said disease is cancer.

14. The method of 1 wherein said tenascin-C nucleic acid ligand is identified by:

i) contacting a candidate mixture of nucleic acids with tenascin-C wherein nucleic acids having an increased affinity to tenascin-C relative to the candidate mixture may be partitioned from the remainder of the candidate mixture;

ii) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture;

iii) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids with relatively higher affinity and specificity for binding to tenascin-C, whereby a nucleic acid ligand of tenascin-C is identified.