Platelet derived growth factor (PDGF) nucleic acid ligand complexes

• US 6,229,002 B1
• Filed: 12/16/1997
• Issued: 05/08/2001
• Est. Priority Date: 06/07/1995
• Status: Expired due to Term

• Current Assignee: Gilead Sciences Inc.
• Original Assignee: NeXstar Pharmaceuticals, Inc. (Gilead Sciences Inc.)
• Inventors: Janjic, Nebojsa, Gold, Larry
• Primary Examiner(s): ZITOMER, STEPHANIE W
• Application Number:
  US08/991,743
• Time in Patent Office: 1,239 Days
• Field of Search: 435/6, 435/91.2, 514/44, 536/24.3, 536/25.4, 536/23.1, 935/72, 935/28
• US Class Current: 536/23.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• 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 47/544 Phospholipids
• CPC Class Codes: A61K 47/60 the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• 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: C07K 14/52 Cytokines; Lymphokines; Interferons
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C12N 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• 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 2310/317 with an inverted bond, e.g. a cap structure
• CPC Class Codes: C12N 2310/3183 Diol linkers, e.g. glycols or propanediols
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3515 : Lipophilic moiety, e.g. cholesterol
• CPC Class Codes: C12N 2310/3521 : Methyl
• CPC Class Codes: C12Q 1/6811 : Selection methods for production or design of target specific oligonucleotides or binding molecules
• CPC Class Codes: C40B 30/04 : by measuring the ability to specifically bind a target molecule, e.g. antibody-antigen binding, receptor-ligand binding

Abstract

This invention discloses a method for preparing a complex comprised of a PDGF Nucleic Acid Ligand and a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound by identifying a PDGF Nucleic Acid Ligand by SELEX methodology and associating the PDGF Nucleic Acid Ligand with a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound. The invention further discloses Complexes comprising one or more PDGF Nucleic Acid Ligands in association with a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound. The invention further includes a Lipid construct comprising a PDGF Nucleic Acid Ligand or Complex and methods for making the same.

Claims

1. A Complex comprised of a platelet-derived growth factor (PDGF) Nucleic Acid Ligand and a Non-Immunogenic, High Molecular Weight Compound.

2. The Complex of claim 1 further comprising a Linker between said Ligand and said Non-Immunogenic, High Molecular Weight Compound.

3. The Complex of claim 1 wherein said Ligand comprises a Linker.

4. The Complex of claim 1 wherein said Non-Immunogenic, High Molecular Weight Compound is a Polyalkylene Glycol.

5. The Complex of claim 4 wherein said Polyalkylene Glycol is polyethylene glycol.

6. The Complex of claim 5 wherein said polyethylene glycol has a molecular weight of about 10-80 K.

7. The Complex of claim 6 wherein said polyethylene glycol has a molecular weight of about 20-45 K.

8. A Lipid Construct comprising the Complex of claim 1.

9. The Complex of claim 1 wherein said PDGF Nucleic Acid Ligand is identified from a Candidate Mixture of Nucleic Acids according to the method comprising:

a) contacting the Candidate Mixture with PDGF, wherein the Nucleic Acids having an increased affinity to PDGF relative to the Candidate Mixture may be partitioned from the remainder of the Candidate Mixture;

b) partitioning the increased affinity Nucleic Acids from the remainder of the Candidate Mixture; and

c) amplifying the increased affinity Nucleic Acids to yield a mixture of Nucleic Acids enriched for Nucleic Acids having an increased affinity for PDGF;

whereby a Nucleic Acid Ligand of PDGF is identified.

10. The Complex of claim 9 wherein the method further comprises repeating steps b) and c).

11. The complex of claim 7 wherein said Complex is

in which said PEG spacer is

and said nucleic acid ligand comprises SEQ ID NO:

146.