Multimeric oligonucleotides having decreased kidney clearance
The present invention relates to methods of administering to a subject multimeric oligonucleotides having monomeric subunits joined by linkers. The multimeric oligonucleotides have a molecular weight of at least about 45 kD and other characteristics, such that their clearance due to glomerular filtration is reduced. The present invention also relates to such multimeric oligonucleotides and methods of synthesizing such multimeric oligonucleotides.
1. A method of administering a multimeric oligonucleotide to a subject in need thereof, the method comprising administering an effective amount of the multimeric oligonucleotide to the subject, the multimeric oligonucleotide comprising subunits , wherein:
each of the subunits is independently a single or double stranded oligonucleotide, and each of the subunits is joined to another subunit by a covalent linker ●;
the multimeric oligonucleotide has a molecular weight and/or size configured to decrease its clearance due to glomerular filtration; and
the molecular weight of the multimeric oligonucleotide is at least about 45 kD; and the decreased clearance due to glomerular filtration results in increased in vivo circulation half-life of the multimeric oligonucleotide.
2. The method of claim 1 , wherein the number of subunits contained in the multimeric oligonucleotide is m, m being an integer selected to enable the multimeric oligonucleotide to have the molecular weight and/or size configured to decrease its clearance due to glomerular filtration.
3. The method of claim 1 , in which the multimeric oligonucleotide comprises Structure 21:
and n is an integer ≥0.
4. The method of claim 1 , in which the subunits are single-stranded oligonucleotides.
5. The method of claim 3 , wherein n is ≥1.
6. The method of claim 1 , in which the subunits are double-stranded oligonucleotides.
7. The method of claim 3 , wherein:
when n=0, the clearance of the multimeric oligonucleotide due to glomerular filtration is decreased relative to that of a monomeric subunit and/or a dimeric subunit
of the multimeric oligonucleotide; and
when n ≥1, the clearance of the multimeric oligonucleotide due to glomerular filtration is decreased relative to that of a monomeric subunit , a dimeric subunit
and/or a trimeric subunit
of the multimeric oligonucleotide.
8. The method of claim 1 , in which the decreased clearance due to glomerular filtration is determined by measuring the in vivo circulation half-life of the multimeric oligonucleotide after administering the multimeric oligonucleotide to the subject.
9. The method of claim 1 , in which the decreased clearance due to glomerular filtration is determined by measuring the time required for the serum concentration of the multimeric oligonucleotide to decrease to a predetermined value.
10. The method of claim 1 , in which the decreased clearance due to glomerular filtration is determined by measuring the serum concentration of the multimeric oligonucleotide at a predetermined time after administering the multimeric oligonucleotide to the subject.
11. The method of claim 1 , in which the decreased clearance due to glomerular filtration is determined by measuring the area under a curve of a graph representing serum concentration of the multimeric oligonucleotide over time after administering the multimeric oligonucleotide to the subject.
12. The method of claim 1 , in which the decreased clearance due to glomerular filtration increases in vivo bioavailability of the multimeric oligonucleotide.
13. The method of claim 1 , in which the decreased clearance due to glomerular filtration increases in vivo cellular uptake of the multimeric oligonucleotide.
14. The method of claim 1 , in which the decreased clearance due to glomerular filtration increases in vivo therapeutic index/ratio of the multimeric oligonucleotide.
15. The method of claim 3 , wherein n is an integer greater than or equal to 1, and at least one of the monomeric subunits comprises a single strand having one of the covalent linkers ● joined to its 3′ terminus and another of the covalent linkers joined to its 5′ terminus.
16. The method of claim 2 , wherein m is greater than or equal to 2.
17. The method of claim 2 , wherein m is greater than or equal to 3.
18. The method of claim 2 , wherein m is greater than or equal to 4.
19. The method of claim 2 , wherein m is 4, 5, 6, 7, or 8.
20. The method of claim 2 , wherein m is greater than or equal to 4 and less than or equal to 17.
21. The method of claim 2 , wherein m is greater than or equal to 4 and less than or equal to 8.
22. The method of claim 1 , wherein the multimeric oligonucleotide further comprises one or more targeting ligands.
23. The method of claim 1 , wherein at least one of the subunits is a targeting ligand.
24. The method of claim 22 , wherein the targeting ligand is an aptamer.
25. The method of claim 22 , wherein the targeting ligand comprises N-Acetylgalactosamine (GalNAc), cholesterol, tocopherol, folate, 2-[3-(1,3-dicarboxypropyl)-ureido]pentanedioic acid (DUPA), or anisamide.
26. The method of claim 22 , wherein the targeting ligand comprises a sugar, peptide, or other nucleic acid.
27. The method of claim 22 , wherein the targeting ligand comprises a cell specific peptide or protein, aptamer, cell growth factor, vitamin, monosaccharide, polysaccharide, arginine-glycine-aspartic acid (RGD), or asialoglycoprotein receptor ligand.
28. The method of claim 27 , wherein the targeting ligand is transferrin.
29. The method of claim 27 , wherein the targeting ligand is folic acid.
30. The method of claim 27 , wherein the targeting ligand is GalNAc.
31. The method of claim 1 , wherein the multimeric oligonucleotide is conjugated to a biologically active moiety that is a protein, peptide, amino acid, other nucleic acid, targeting ligand, carbohydrate, polysaccharide, lipid, other organic compound, or inorganic compound.
32. The method of claim 31 , wherein the biologically active moiety has gene editing activity.
33. The method of claim 32 , wherein the biologically active moiety having gene editing activity is an oligonucleotide.
34. The method of claim 1 , wherein the multimeric oligonucleotide comprises Structure 54:
wherein each subunit is independently a double-stranded oligonucleotide , wherein d is an integer greater than or equal to 1, and wherein each covalent linker ● is on the same strand.
35. The method of claim 1 , wherein the multimeric oligonucleotide comprises Structure 28:
36. The method of claim 1 , wherein the multimeric oligonucleotide comprises Structure 30:
37. The method of claim 1 , wherein the multimeric oligonucleotide does not comprise PEG.
38. The method of claim 1 , comprising two or more different covalent linkers.
39. The method of claim 1 , wherein at least one of the covalent linkers ● joins three or more monomeric subunits .
40. The method of claim 1 , wherein the multimeric oligonucleotide comprises Structure 29:
41. The method of claim 1 , where the multimeric oligonucleotide comprises
42. The method of claim 2 , wherein each monomeric subunit is independently a double stranded oligonucleotide, and wherein m is 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.
43. The method of claim 2 , wherein each monomeric subunit is independently a double stranded oligonucleotide, wherein m is 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12, and wherein each covalent linker is on the same strand.
44. The method of claim 1 , where the multimeric oligonucleotide comprises