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Claims for Patent: 6,147,204

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Claims for Patent: 6,147,204

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 (Niwot, CO), Janjic; Nebojsa (Boulder, CO)
Assignee: NeXstar Pharmaceuticals, Inc. (Boulder, CO)
Application Number:08/945,604
Patent Claims: 1. A therapeutic or diagnostic Complex comprised of a Nucleic Acid Ligand and a Lipophilic Compound or a Non-Immunogenic, High Molecular Weight Compound, wherein said Nucleic Acid Ligand is covalently linked to said Lipophilic Compound or said Non-Immunogenic, High Molecular Weight Compound, and wherein said Nucleic Acid Ligand has a specific binding affinity for a target molecule, such 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 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 was identified as a ligand of a given Target from a Candidate Mixture of Nucleic Acids according to the method comprising:

a) contacting the Candidate Mixture with the Target, wherein the Nucleic Acids having an increased affinity to the Target 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 are identified.

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

d) repeating steps b) and c).

4. The Complex of claim 1 comprised of a Nucleic Acid Ligand and a Non-Immunogenic, High Molecular Weight Compound.

5. The Complex of claim 4 wherein said Non-Immunogenic, High Molecular Weight Compound is selected from the group consisting of polyethylene glycol, dextran, albumin, and magnetite.

6. The Complex of claim 4 wherein said Non-Immunogenic, High Molecular Weight Compound is polyethylene glycol.

7. The Complex of claim 1 comprised of a Nucleic Acid Ligand and a Lipophilic Compound.

8. The Complex of claim 7 wherein said Lipophilic Compound is selected from the group consisting of cholesterol, dialkyl glycerol, and diacyl glycerol.

9. The Complex of claim 8 wherein said Lipophilic Compound is cholesterol.

10. The Complex of claim 8 wherein said Lipophilic Compound is dialkyl glycerol.

11. The Complex of claim 9 further comprising a Lipid Construct.

12. The Complex of claim 11 wherein said Lipid Construct is a Lipid Bilayer Vesicle.

13. The Complex of claim 12 wherein said Lipid Bilayer Vesicle is a Liposome.

14. The Complex of claim 7 further comprising a Lipid Construct.

15. The Complex of claim 14 wherein said Lipid Construct is a Lipid Bilayer Vesicle.

16. The Complex of claim 15 wherein said Lipid Bilayer Vesicle is a Liposome.

17. The Complex of claim 10 further comprising a Lipid Construct.

18. The Complex of claim 17 wherein said Lipid Construct is a Lipid Bilayer Vesicle.

19. The Complex of claim 18 wherein said Lipid Construct is a Liposome.

20. The Complex of claim 6 further comprising a Lipid Construct.

21. The Complex of claim 20 wherein said Lipid Construct is a Lipid Bilayer Vesicle.

22. The Complex of claim 21 wherein said Lipid Bilayer Vesicle is a Liposome.

23. The Complex of claim 16 wherein said Nucleic Acid Ligand is encapsulated within the interior of said Liposome.

24. The Complex of claim 16 wherein said Nucleic Acid Ligand protrudes from the exterior of said Liposome.

25. The Complex of claim 16 wherein said Nucleic Acid Ligand is covalently attached to a second Lipophilic Compound.

26. The Complex of claim 25 wherein said Nucleic Acid Ligand is attached to a second Lipophilic Compound via a Linker.

27. The Complex of claim 25 wherein said Nucleic Acid Ligand projects out of the exterior surface of said Liposome.

28. The Complex of claim 25 wherein said Nucleic Acid Ligand projects into the interior of said Liposome.

29. The Complex of claim 16 further comprising and additional therapeutic or diagnostic agent.

30. The Complex of claim 29 wherein said therapeutic agent is encapsulated in the interior of said Liposome.

31. The Complex of claim 29 wherein said additional therapeutic or diagnostic agent protrudes from the membrane of said Liposome.

32. The Complex of claim 29 wherein said additional therapeutic or diagnostic agent protrudes from the exterior surface of said Liposome.

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

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

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

36. The Complex of claim 35 wherein the cellular uptake of said Nucleic Acid Ligand is enhanced relative to the Nucleic Acid Ligand alone.

37. The Complex of claim 6 further comprising an additional therapeutic or diagnostic agent.

38. The Complex of claim 37 wherein said addition therapeutic or diagnostic agent is covalently or noncovalently linked to said polyethylene glycol.

39. The Complex of claim 38 wherein said additional therapeutic or diagnostic agent is covalently linked to said polyethylene glycol.
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