Modulators of pharmacological agents
The biological activity of nucleic acid ligand is regulated (i.e. enhanced or inhibited) in vivo to produce a desired biological effect. This is accomplished through the administration of a modulator, or regulator, that changes the binding of the nucleic acid ligand for its target or that degrades or otherwise cleaves, metabolizes or breaks down the nucleic acid ligand while the ligand is still exerting its effect. Modulators of the present invention can be administered in real time as needed based on various factors, including the progress of the patient, as well as the physician's discretion in how to achieve optimal therapy. Thus, this invention provides for the first time a regulatable therapeutic regime in the course of nucleic acid ligand therapy.
1. A method of altering the affinity of a nucleic acid ligand for a coagulation factor in a mammalian host comprising administering an oligonucleotide nucleic acid modulator to the host receiving the nucleic acid ligand under conditions such that the modulator hybridizes to the nucleic acid ligand and thereby alters the affinity of the nucleic acid ligand for the coagulation factor wherein the affinity of the nucleic acid ligand for the coagulation factor is reduced.
2. The method according to claim 1 wherein the modulator binds to free nucleic acid ligand present in the host.
3. The method according to claim 1 wherein the modulator binds to nucleic acid ligand present in the host in association with the coagulation factor.
4. The method according to claim 1 wherein the nucleic acid modulator is an oligonucleotide complementary to the nucleic acid ligand.
5. The method according to claim 4 wherein the oligonucleotide comprises a sequence complementary to 6-25 nucleotides of the nucleic acid ligand.
6. The method according to claim 5 wherein the oligonucleotide comprises a sequence complementary to 8-20 nucleotides of the nucleic acid ligand.
7. The method according to claim 6 wherein the oligonucleotide comprises a sequence complementary to 10-15 nucleotides of the nucleic acid ligand.
8. The method according to claim 4 wherein the oligonucleotide comprises 5-80 nucleotides.
9. The method according to claim 8 wherein the oligonucleotide comprises 10-30 nucleotides.
10. The method according to claim 9 wherein the oligonucleotide comprises 15-20 nucleotides.
11. The method according to claim 4 wherein the oligonucleotide bears a chemical substitution.
12. The method according to claim 11 wherein the oligonucleotide comprises a pyrimidine substituted at a 5 position or a sugar substituted at a 2′ position.
13. The method according to claim 12 wherein the oligonucleotide comprises a 2′-amino, 2′-fluoro or 2′-O-methyl substitution.
14. The method according to claim 4 wherein the oligonucleotide comprises a locked nucleic acid.
15. The method according to claim 4 wherein the oligonucleotide is complementary to a single-stranded region of said nucleic acid ligand.
16. The method according to claim 15 wherein the oligonucleotide is complementary to a single-stranded region of the nucleic acid ligand and a double stranded region of the nucleic acid ligand.
17. The method according to claim 4 wherein the nucleic acid ligand comprises a single-stranded tail.
18. The method according to claim 17 wherein the oligonucleotide is complementary to the single-stranded tail of the nucleic acid ligand.
19. The method according to claim 4 wherein the nucleic acid ligand and the oligonucleotide comprise β-D-nucleotides.
20. The method according to claim 4 wherein the oligonucleotide is produced in the host following administration of a construct comprising a sequence encoding the oligonucleotide to the host.
21. The method according to claim 1 wherein the nucleic acid ligand is an anticoagulant or antithrombotic nucleic acid ligand.
22. The method according to claim 21 wherein the modulator reverses the anticoagulant or antithrombotic effect of the nucleic acid ligand.
23. The method according to claim 21 wherein the nucleic acid ligand binds to a tissue factor (TF)/factor VIIa (FVIIa) enzyme complex, factor VIIIa (FVIIIa)/factor IXa (FIXa) enzyme complex, factor Va (FVa)/factor Xa (FXa) enzyme complex, gpIIbIIIa, gpIbIX, gpVI, Gas6, PAI-1 (plasminogen activator inhibitor 1), coagulation factor XIIIa (FXIIIa), ATIII (anti-thrombin III), coagulation factor IXa (FIXa), thrombin or coagulation factor XIa (FXIa).
24. A method of reversing binding of a nucleic acid ligand bearing a label to a coagulation factor on a target tissue comprising administering to a mammalian host receiving the nucleic acid ligand an oligonucleotide nucleic acid modulator that hybridizes to the nucleic acid ligand and inhibits the binding of the nucleic acid ligand to the coagulation factor on the target tissue.
25. The method according to claim 24 wherein the modulator is an oligonucleotide complementary to the nucleic acid ligand.
26. The method according to claim 24 wherein the label is a cytotoxic label.
27. The method according to claim 24 wherein the label is a radioactive label.
28. The method according to claim 24 wherein the label is a detectable label.
29. The method according to claim 1 or 24 wherein the host is a human.
30. The method according to claim 1 or 24 wherein the host is a non-human mammal.
31. The method of claim 21 wherein the nucleic acid ligand binds to a factor VIIIa (FVIIIa)/factor IXa (FIXa) complex.
32. The method of claim 21 wherein the nucleic acid ligand binds to Factor IXa.