High affinity ligands of basic fibroblast growth factor and thrombin

• US 6,177,557 B1
• Filed: 08/05/1996
• Issued: 01/23/2001
• Est. Priority Date: 06/11/1990
• Status: Expired due to Fees

• Current Assignee: Gilead Sciences Inc., The University Court Of The University Of Glasgow
• Original Assignee: NeXstar Pharmaceuticals, Inc. (Gilead Sciences Inc.)
• Inventors: Tasset, Diane, Janjic, Nebojsa, Gold, Larry
• Primary Examiner(s): Zitomer, Stephanie W.
• Application Number:
  US08/687,421
• Time in Patent Office: 1,632 Days
• Field of Search: 435/6, 435/91.2, 536/23.1, 536/24.31, 536/25.4, 514/44, 935/72, 935/78
• US Class Current: 536/24.31
• 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: 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: C12Q 1/6811 Selection methods for production or design of target specific oligonucleotides or binding molecules
• CPC Class Codes: C40B 40/00 Libraries per se, e.g. arrays, mixtures

Abstract

The present invention utilizes the SELEX (Systematic Evolution of Ligands for EXponential Enrichment) method for identifying and preparing nucleic acid ligands to basic fibroblast growth factor (bFGF) and thrombin. Included in the invention are nucleic acid ligands to bFGF which are inhibitors of bFGF and 2′-amino-modified RNA ligands to bFGF. Further included in the present invention are modified nucleotide sequences to thrombin based on the sequences of the RNA ligands identified. The modified RNA ligands to bFGF and thrombin exhibit increased in vivo stability.

Claims

1. A purified and isolated non-naturally occurring DNA ligand to bFGF wherein the nucleic acid sequence of said ligand is selected from the group consisting of SEQ ID NOS:

330-444.

2. A purified and isolated non-naturally occurring nucleic acid ligand to bFGF wherein the nucleic acid sequence of said ligand is substantially homologous to and has substantially the same ability to bind bFGF as a ligand selected from the group consisting of SEQ ID NOS:

7-69, 101-185, 281-319 and 330-444.

3. A purified and isolated non-naturally occurring nucleic acid ligand to bFGF wherein said ligand has substantially the same structure and substantially the same ability to bind bFGF as a ligand selected from the group consisting of SEQ ID NOS:

7-69, 101-185, 281-319 and 330-444.