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Claims for Patent: 6,011,020

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Claims for Patent: 6,011,020

Title: Nucleic acid ligand complexes
Abstract:This invention discloses a method for preparing a therapeutic or diagnostic complex comprised of a nucleic acid ligand and a lipophilic compound or non-immunogenic, high molecular weight compound by identifying a nucleic acid ligand by SELEX methodology and associating the nucleic acid ligand with a lipophilic compound or a non-immunogenic, high molecular weight compound. The invention further discloses complexes comprising one or more nucleic acid ligands in association with a lipophilic compound or non-immunogenic, high molecular weight compound.
Inventor(s): Gold; Larry (Boulder, CO), Schmidt; Paul G. (San Marino, CA), Janjic; Nebojsa (Boulder, CO)
Assignee: NeXstar Pharmaceuticals, Inc. (Boulder, CO)
Application Number:08/434,465
Patent Claims: 1. A therapeutic or diagnostic Complex comprised of a Nucleic Acid Ligand and a polyethylene glycol (PEG), wherein said Nucleic Acid Ligand is covalently linked to said PEG, and wherein said Nucleic Acid Ligand has a specific binding affinity for a Target Molecule, said Target Molecule being a three dimensional chemical structure other than a polynucleotide that binds to said Nucleic Acid Ligand through a mechanism which predominantly depends on Watson/Crick base pairing or triple helix binding, wherein said Nucleic Acid Ligand is not a Nucleic Acid having the known physiological function of being bound by the Target Molecule, wherein said PEG is at least about 1000 Daltons, and wherein said Complex has improved pharmacokinetic properties relative to the pharmacokinetic properties of the Nucleic Acid Ligand alone.

2. The Complex of claim 1 wherein said Nucleic Acid Ligand is identified as a ligand of a Target Molecule from a Candidate Mixture of Nucleic Acids according to the method comprising:

a) contacting the Candidate Mixture with the Target Molecule, wherein the Nucleic Acids having an increased affinity to the Target Molecule 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 ligand-enriched mixture of Nucleic Acids, whereby a Nucleic Acid Ligand of said Target Molecule is identified.

3. The Complex of claim 2 wherein said method further comprises:

d) repeating steps b) and c).

4. The Complex of claim 1 comprising a Lipid Construct.

5. The Complex of claim 4 wherein said Lipid Construct is a Lipid Bilayer Vesicle.

6. The Complex of claim 5 wherein said Lipid Bilayer Vesicle is a Liposome.

7. The Complex of claim 1 wherein said Nucleic Acid Ligand targets the Complex to a preselected location.

8. The Complex of claim 1 wherein the Target Molecule of said Nucleic Acid Ligand is an intercellular Target Molecule.

9. The Complex of claim 1 wherein the Target Molecule of said Nucleic Acid Ligand is an intracellular Target Molecule.

10. The Complex of claim 9 wherein said Nucleic Acid Ligand has enhanced cellular uptake relative to the Nucleic Acid Ligand alone.

11. The Complex of claim 1 further comprising an additional therapeutic or diagnostic agent.

12. The Complex of claim 11 wherein said additional therapeutic or diagnostic agent is associated with said polyethylene glycol.

13. The Complex of claim 12 wherein said additional therapeutic or diagnostic agent is covalently associated with said polyethylene glycol.

14. A method for improving the pharmacokinetic properties of a Nucleic Acid Ligand comprising covalently linking said Nucleic Acid Ligand to a PEG, wherein said Nucleic Acid Ligand has a specific binding affinity for a Target Molecule, said Target Molecule being a three dimensional chemical structure other than a polynucleotide that binds to said Nucleic Acid Ligand through a mechanism which predominantly depends on Watson/Crick base pairing or triple helix binding, wherein said Nucleic Acid Ligand is not a Nucleic Acid having the known physiological function of being bound by the Target Molecule, and wherein said PEG is at least about 1000 Daltons.

15. The method of claim 14 wherein said Nucleic Acid Ligand is identified from a Candidate Mixture of Nucleic Acids, said Nucleic Acid Ligand being a ligand of said Target Molecule, by the method comprising:

a) contacting the Candidate Mixture with the Target Molecule, wherein Nucleic Acids having an increased affinity to the Target Molecule 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 ligand-enriched mixture of Nucleic Acids, whereby Nucleic Acid Ligands of said Target Molecule are identified.

16. The method of claim 14 wherein said Nucleic Acid Ligand is linked to a Lipid Construct.

17. The method of claim 16 wherein said Lipid Construct is a Lipid Bilayer Vesicle.

18. The method of claim 17 wherein said Lipid Bilayer Vesicle is a Liposome.

19. The method of claim 14 wherein said Nucleic Acid Ligand targets to a preselected location.

20. The method of claim 15 wherein said Target Molecule is an intercellular Target Molecule.

21. The method of claim 15 wherein said Target Molecule is an intracellular Target Molecule.

22. The method of claim 14 which further comprises attaching an additional therapeutic or diagnostic agent with said Nucleic Acid Ligand.

23. The method of claim 22 wherein said additional therapeutic or diagnostic agent is attached to said PEG.

24. The method of claim 23 wherein said additional therapeutic or diagnostic agent is covalently attached to said PEG.
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