Vascular endothelial growth factor (VEGF) nucleic acid ligand complexes

• US 6,426,335 B1
• Filed: 03/13/2000
• Issued: 07/30/2002
• Est. Priority Date: 10/17/1997
• Status: Expired due to Term

• Current Assignee: Gilead Sciences Inc.
• Original Assignee: Gilead Sciences Inc.
• Inventors: Vargeese, Chandra, Willis, Michael, Janjic, Nebojsa, Gold, Larry, Schmidt, Paul
• Primary Examiner(s): ZITOMER, STEPHANIE W
• Application Number:
  US09/254,968
• Time in Patent Office: 869 Days
• Field of Search: 514/44, 514/23.1, 536/23.1
• US Class Current: 514/44.00R
• CPC Class Codes: A61K 31/70 Carbohydrates; Sugars; Derivatives thereof sorbitol A61K31/047
• 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: 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 14/52 Cytokines; Lymphokines; Interferons
• 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 15/88 using microencapsulation, e.g. using amphiphile liposome vesicle
• CPC Class Codes: C12N 2310/3515 Lipophilic moiety, e.g. cholesterol

Abstract

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

Claims

1. A method for treating a VEGF mediated disease or medical condition selected from the group consisting of cancer, psoriasis, ocular disorders characterized by excessive angiogenesis, collagen vascular diseases and rheumatoid arthritis comprising administering a pharmaceutically effective amount of a Complex comprised of a VEGF Nucleic Acid Ligand comprising 2′

F-modified nucleotides and a Non-Immunogenic, High Molecular Weight Compound or a Lipophilic Compound to a patient in need of said Complex.

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

3. The method of claim 2 wherein said Polyalkylene Glycol is polyethylene glycol.

4. The method of claim 3 wherein said polyethylene glycol has a molecular weight of about between 10-80K.

5. The method of claim 3 wherein said polyethylene glycol has a molecular weight of about 20-45K.

6. The method of claim 5 wherein said Complex has the structure

Ligand Component=fCmGmGrArAfUfCmAmGfUmGmAmAfUmGfCfUfUmAfUmAfCmAfUfCfCmG-3′

3′

-dT (VEGF ligand).

7. The method of claim 1 wherein said Lipophilic Compound is a glycerol lipid.

8. The method of claim 7 wherein said Complex is further associated with a lipid construct.

9. The method of claim 8 wherein said Complex has the structure

Ligand Component=fCmGmGrArAfUfCmAmGfUmGmAmAfUmGfCfUfUmAfUmAfCmAfUfCfCmG-3′

3′

-dT (VEGF ligand).

10. A method for improving the pharmacokinetic properties of a VEGF Nucleic Acid Ligand comprising 2′

F-modified nucleotides comprising;

covalently linking said VEGF Nucleic Acid Ligand with a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound to form a Complex comprised of a VEGF Nucleic Acid Ligand and a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound; and

administering said Complex to a patient, whereby the pharmacokinetic properties of said VEGF Nucleic Acid Ligand are improved.

11. A method for targeting a therapeutic or diagnostic agent to a specific predetermined biological target that is expressing VEGF in a patient comprising:

covalently linking said therapeutic or diagnostic agent with a Complex comprised of a VEGF Nucleic Acid Ligand comprising 2′

F-modified nucleotides and a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound, and administering said Complex to said patient, whereby said therapeutic or diagnostic agent is targeted to a specific predetermined biological target that is expressing VEGF.

12. A method of inhibiting VEGF mediated angiogenesis in a patient comprising:

covalently linking a VEGF Nucleic Acid Ligand comprising 2′

F-modified nucleotides to a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound to form a Complex, said Complex being capable of inhibiting VEGF mediated angiogenesis; and

administering said Complex to said patient, whereby said VEGF mediated angiogenesis is inhibited.

13. A method of inhibiting the growth of tumors in a patient comprising:

covalently linking a VEGF Nucleic Acid Ligand comprising 2′

F-modified nucleotides to a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound to form a Complex, said Complex being capable of inhibiting the growth of tumors; and

administering said Complex to said patient, whereby said growth of tumors is inhibited.

14. The method of claim 13 wherein said tumors or cells or tissues surrounding said tumors are expressing VEGF.

15. The method of claim 13 wherein said tumors or cells or tissues surrounding said tumors are expressing VEGF receptors.

16. The method of claim 15 wherein said tumors are comprised of Kaposi'"'"'s sarcoma cells.

17. A method of inhibiting macular degeneration in a patient comprising:

covalently linking a VEGF Nucleic Acid Ligand comprising 2′

F-modified nucleotides to a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound to form a Complex, said Complex being capable of inhibiting macular degeneration; and

administering said Complex to said patient, whereby said macular degeneration is inhibited.

18. The method of claim 17 wherein said Non-Immunogenic, High Molecular Weight Compound is a polyalkylene glycol.

19. The method of claim 18 wherein said polyalkylene glycol is polyethylene glycol.

20. The method of claim 19 wherein said polyethylene glycol has a molecular weight of about between 10-80k.

21. The method of claim 19 wherein said polyethylene glycol has a molecular weight of about between 20-45k.

22. The method of claim 21 wherein said complex has the structure

Ligand Component=fCmGmGrArAfUfCmAmGfUmGmAmAfUmGfCfUfUmAfUmAfCmAfUfCfCmG-3′ 3′

-dT (VEGF ligand).

23. A method for prolonging the residence time of a Nucleic Acid Ligand in an ocular application comprising attaching a Non-Immunogenic, High Molecular Weight Compound to a Nucleic Acid Ligand to form a Complex, and administering said complex to the eye, whereby the residence time of said Nucleic Acid Ligand is prolonged.