High affinity oligonucleotide ligands to growth factors

• US 6,207,816 B1
• Filed: 05/11/1998
• Issued: 03/27/2001
• Est. Priority Date: 06/02/1995
• Status: Expired due to Term

• Current Assignee: Gilead Sciences Inc.
• Original Assignee: NeXstar Pharmaceuticals, Inc. (Gilead Sciences Inc.)
• Inventors: Janjic, Neboisa, Gold, Larry, Pagratis, Nikos
• Primary Examiner(s): ZITOMER, STEPHANIE W
• Application Number:
  US08/973,124
• Time in Patent Office: 1,051 Days
• Field of Search: 435/6, 435/91.2, 536/24.31, 536/25.4, 935/77, 935/78
• US Class Current: 536/24.1
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C07K 14/475 Growth factors; Growth regulators
• CPC Class Codes: C07K 14/49 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C07K 14/495 Transforming growth factor [TGF]
• CPC Class Codes: C07K 14/50 Fibroblast growth factor [FGF]
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding 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: C12N 2320/13 in a process of directed evolution, e.g. SELEX, acquiring a new function
• CPC Class Codes: G01N 2333/96433 Serine endopeptidases (3.4.21)

Abstract

Methods are described for the identification and preparation of high-affinity nucleic acid ligands to TGFβ, PDGF and hKGF. Included in the invention are specific RNA and ssDNA ligands to TGFβ1 and PDGF identified by the SELEX method. Also included in the invention are specific RNA ligands to hKGF identified by the SELEX method. Further included are RNA ligands that inhibit the interaction of TGFβ1 and hKGF with their receptors and DNA ligands that inhibit the interaction of PDGF with its receptor.

Claims

1. A purified and isolated non-naturally occurring modified single-stranded ribonucleic acid ligand to platelet-derived growth factor (PDGF) wherein said modified ligand is substantially homologous to and has substantially the same ability to bind PDGF as a modified ligand selected from the group consisting of the sequences set forth in Table 13 (SEQ ID NOS:

128-170).

2. A purified and isolated non-naturally occurring modified single-stranded ribonucleic acid ligand to platelet-derived growth factor (PDGF) wherein said modified ligand has substantially the same structure and substantially the same ability to bind PDGF as a modified ligand selected from the group consisting of the sequences set forth in Table 13 (SEQ ID NOS:

128-170).

3. A purified and isolated non-naturally occurring single-stranded deoxyribonucleic acid ligand to platelet-derived growth factor (PDGF) wherein said ligand is substantially homologous to and has substantially the same ability to bind PDGF as a ligand selected from the group consisting of the sequences set forth in Tables 8 and 9, and FIGS. 3, 4 and 9 (SEQ ID NOS:

93-124, 171-176).

4. A purified and isolated non-naturally occurring single stranded deoxyribonucleic acid ligand to platelet-derived growth factor (PDGF) wherein said ligand has substantially the same structure and substantially the same ability to bind PDGF as a ligand selected from the group consisting of the sequences set forth in Tables 8 and 9, and FIGS. 3, 4 and 9 (SEQ ID NOS:

93-124, 171-176).

5. A purified and isolated non-naturally occurring modified single-stranded ribonucleic acid ligand to human keratinocyte growth factor (hKGF) wherein said modified ligand is selected from the group consisting of SEQ ID NOS:

189-264, 268-304.

6. A purified and isolated non-naturally occurring modified single-stranded ribonucleic acid ligand to human keratinocyte growth factor (hKGF) wherein said modified ligand is substantially homologous to and has substantially the same ability to bind hKGF as a modified ligand selected from the group consisting of SEQ ID NOS:

189-264, 268-304.

7. A purified and isolated non-naturally occurring modified ribonucleic acid ligand to human keratinocyte growth factor (hKGF) wherein said modified ligand has substantially the same structure and substantially the same ability to bind hKGF as a modified ligand selected from the group consisting of SEQ ID NOS:

189-264, 268-304.

8. A purified and isolated non-naturally occurring modified ribonucleic acid ligand to basic fibroblast growth factor (bFGF) wherein said modified ligand has the sequence as shown in SEQ ID NO:

267.