Methods for Oral Delivery of Oligonucleotides
Provided herein are methods for oral administration of oligonucleotides. Further provided herein are methods for oral administration of modified oligonucleotides targeted to microRNA.
1 . A method of inhibiting the activity of a microRNA, comprising administering to a subject a compound comprising a modified oligonucleotide complementary to the microRNA, or a pharmaceutically acceptable salt thereof, wherein the modified oligonucleotide has a length of 6 to 25 linked nucleotides and wherein the administration is oral administration.
2 . The method of claim 1 , wherein the modified oligonucleotide is fully complementary to the microRNA.
3 . The method of claim 1 or 2 , wherein the microRNA is expressed in the kidney and the compound consists of the modified oligonucleotide.
4 . The method of claim 1 or 2 , wherein the microRNA is expressed in the liver and the compound comprises the modified oligonucleotide linked to a conjugate moiety.
5 . The method of any one of claims 1 to 4 , wherein the modified oligonucleotide is 8 to 13 linked nucleosides in length, or 8 to 12 linked nucleotides in length.
6 . The method of any one of claims 1 to 4 , wherein the modified oligonucleotide is 8 linked nucleosides in length.
7 . The method of any one of claims 1 to 4 , wherein the modified oligonucleotide is 9 linked nucleosides in length.
8 . The method of any one of claims 1 to 4 , wherein the modified oligonucleotide is 10 linked nucleosides in length.
9 . The method of any one of claims 1 to 4 , wherein the modified oligonucleotide is 11 linked nucleosides in length.
10 . The method of any one of claims 1 to 4 , wherein the modified oligonucleotide is 12 linked nucleosides in length.
11 . The method of any one of claims 1 to 4 , wherein the modified oligonucleotide is 13 linked nucleosides in length.
12 . The method of any one of claims 1 to 11 , wherein the modified oligonucleotide comprises at least one nucleoside with a modified sugar moiety.
13 . The method of claim 12 , wherein each nucleoside of the modified oligonucleotide comprises a modified sugar moiety.
14 . The method of claim 12 or 13 , wherein each modified sugar moiety is independently selected from a 2′-O-methyl sugar moiety, a 2′-O-methoxyethyl sugar moiety, a 2′-fluoro sugar moiety, and a bicyclic sugar moiety.
15 . The method of claim 14 , wherein each bicyclic sugar moiety is independently selected from a cEt sugar moiety and an LNA sugar moiety.
16 . The method of claim 15 , wherein the cEt nucleoside is an S-cEt nucleoside.
17 . The method of any of claims 1 to 11 , wherein the modified oligonucleotide comprises a plurality of non-bicyclic nucleosides and a plurality of bicyclic nucleosides.
18 . The method of claim 17 , wherein each non-bicyclic nucleoside is independently selected from a 2′-O-methyl nucleoside, a 2′-O-methoxyethyl nucleoside, and a 2′-fluoronucleoside.
19 . The method of claim 18 , wherein each bicyclic nucleoside is selected from a cEt nucleoside and an LNA nucleoside.
20 . The method of claim 19 , wherein the cEt nucleoside is an S-cEt nucleoside.
21 . The method of any one of claims 1 to 20 , wherein the modified oligonucleotide comprises at least one modified internucleoside linkage.
22 . The method of any one of claims 1 to 20 , wherein each internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage.
23 . The method of claim 21 or 22 , wherein the modified internucleoside linkage is a phosphorothioate linkage.
24 . The method of claim 4 , wherein the conjugate moiety comprises a cholesterol moiety or a carbohydrate moiety.
25 . The method of claim 24 , wherein the carbohydrate moiety is selected from is selected from N-acetylgalactosamine, galactose, galactosamine, N-formylgalactosamine, N-propionyl-galactosamine, N-n-butanoylgalactosamine, and N-iso-butanoyl-galactosamine.
26 . The method of claim 4 , wherein the compound has the structure:
L n -linker-X 1 —N m —X 2 -MO;
wherein each L is, independently, a ligand and n is from 1 to 10; each N is, independently, a modified or unmodified nucleoside and m is from 1 to 5; X 1 is a phosphodiester linkage or a phosphorothioate linkage; X 2 is a phosphodiester linkage or a phosphorothioate linkage; and MO is the modified oligonucleotide.
27 . The method of claim 26 comprising the structure:
wherein:
B is selected from —O—, —S—, —N(R N )—, —Z—P(Z′)(Z″)O—, —Z—P(Z′)(Z″)O—N m —X 1 —, and —Z—P(Z′)(Z″)O—N m —X 2 —;
MO is the modified oligonucleotide;
R N is selected from H, methyl, ethyl, propyl, isopropyl, butyl, and benzyl;
Z, Z′, and Z″ are each independently selected from O and S;
each N is, independently, a modified or unmodified nucleoside;
m is from 1 to 5;
X 1 is selected from a phosphodiester linkage and a phosphorothioate linkage;
X 2 is a phosphodiester linkage; and
the wavy line indicates the connection to the rest of the linker and ligand(s).
28 . The method of claim 26 or 27 , wherein n is from 1 to 5, 1 to 4, 1 to 3, or 1 to 2.
29 . The method of claim 26 or 27 , wherein n is 3 and each ligand is N-acetylgalactosamine.
30 . The method of claim 26 or 27 , wherein n is 1 and the ligand is cholesterol.
31 . The method of claim 26 or 27 , wherein the compound has the structure:
wherein each N is, independently, a modified or unmodified nucleoside and m is from 1 to 5; X 1 and X 2 are each, independently, a phosphodiester linkage or a phosphorothioate linkage; and MO is the modified oligonucleotide.
32 . The method of claim 26 or 27 , wherein the compound has the structure:
wherein each N is, independently, a modified or unmodified nucleoside and m is from 1 to 5; X 1 and X 2 are each, independently, a phosphodiester linkage or a phosphorothioate linkage; and MO is the modified oligonucleotide.
33 . The method of claim 31 or 32 , wherein at least one of X 1 and X 2 is a phosphodiester linkage.
34 . The method of claim 31 or 32 , wherein each of X 1 and X 2 is a phosphodiester linkage.
35 . The method of any one of claims 26 to 34 , wherein m is 1.
36 . The method of any one of claims 26 to 34 , wherein m is 2, 3, 4, or 5.
37 . The method of any one of claims 26 to 36 , wherein N m is N′ p N″, wherein each N′ is, independently, an unmodified nucleoside and p is from 0 to 4; and N″ is a nucleoside comprising an unmodified sugar moiety.
38 . The method of claim 37 , wherein p is 0.
39 . The method of claim 37 or 38 , wherein the unmodified sugar moiety is a β-D-ribose or a β-D-deoxyribose.
40 . The method of claim 39 , wherein the β-D-deoxyribose is β-D-deoxyriboadenosine.
41 . The method of any one of claims 1 to 40 , wherein the compound is present in a pharmaceutical composition.
42 . The method of claim 41 , wherein the pharmaceutical composition comprises a pharmaceutically acceptable diluent.
43 . The method of claim 42 , wherein the pharmaceutically acceptable diluent is an aqueous solution.
44 . The method of claim 43 , wherein the aqueous solution is a saline solution.
45 . The method of claim 43 or 44 , wherein the aqueous solution comprises sodium bicarbonate.