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Details for Patent: 6,962,784

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Details for Patent: 6,962,784

Title: Vascular endothelial growth factor (VEGF) nucleic acid ligand complexes
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.
Inventor(s): Janjic; Nebojsa (Boulder, CO), Gold; Larry (Boulder, CO), Schmidt; Paul (Niwot, CO), Vargeese; Chandra (Thornton, CO), Willis; Michael (Louisville, CO)
Assignee: Gilead Sciences, Inc. (Foster City, CA)
Filing Date:Jul 25, 2002
Application Number:10/205,009
Claims:1. A method for the preparation of 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, said method comprising: a) identifying a VEGF Nucleic Acid Ligand from a Candidate Mixture of Nucleic Acids by the method comprising: i) contacting the Candidate Mixture with VEGF, wherein Nucleic Acids having an increased affinity to VEGF relative to the Candidate Mixture may be partitioned from the remainder of the Candidate Mixture; ii) partitioning the increased affinity VEGF Nucleic Acids from the remainder of the Candidate Mixture; iii) amplifying the increased affinity VEGF Nucleic Acids to yield a ligand-enriched mixture of Nucleic Acids; and b) associating said identified VEGF Nucleic Acid Ligand with a Non-Immunogenic, High Molecular Weight Compound or Lipophilic Compound.

2. The method of claim 1 wherein said Complex is further associated with a Lipid Construct.

3. The method of claim 2 wherein said Lipid Construct is a Liposome.

4. The method of claim 3 wherein said Complex is comprised of a Nucleic Acid Ligand and a Lipophilic Compound and wherein said Complex is passively associated with the bilayer of said Liposomes by the method comprising the steps of: a) forming a liposome; and b) mixing said Complex comprised of a Nucleic Acid Ligand and a Lipophilic Compound with the Liposomes of step a) whereby the Nucleic Acid Ligand Component of said Complex becomes associated with the bilayer of the Liposome and projects from the exterior of the Lipid bilayer.

5. The method of claim 1 wherein said Non-immunogenic, High Molecular Weight Compound is Polyalkylene Glycol.

6. The method of claim 5 wherein said Polyalkylene Glycol is polyethylene glycol.

7. The method of claim 6 wherein said polyethylene glycol has a molecular weight of about between 10-80 K.

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

9. The method of claim 8 wherein said Complex has the structure ##STR10##

(VEGF ligand)

(SEQ. ID. NO. 5).

10. The method of claim 8 wherein said Complex has the structure ##STR11##

(VEGF ligand)

(SEQ. ID. NO. 5).

11. The method of claim 8 wherein said Complex has the structure ##STR12##

(VEGF ligand)

(SEQ. ID. NO. 5).

12. The method of claim 8 wherein said Complex has the structure ##STR13##

(VEGF ligand)

(SEQ. ID. NO. 5).

13. The method of claim 8 wherein said Complex has the structure ##STR14##

(VEGF ligand)

(SEQ. ID. NO. 5).

14. The method of claim 2 wherein said Complex is ##STR15##

(VEGF ligand)

(SEQ. ID. NO. 5).

15. The method of claim 2 wherein said Complex is ##STR16##

(VEGF ligand)

(SEQ. ID. NO. 5).

16. The method of claim 2 wherein said Complex is ##STR17##

(VEGF ligand)

(SEQ. ID. NO. 5).

17. The method of claim 2 wherein said Complex is ##STR18##

(VEGF ligand)

(SEQ. ID. NO. 2).
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