Carbohydrate conjugates as delivery agents for oligonucleotides
View Patent ↗The present invention provides iRNA agents comprising at least one subunit of the formula (I): wherein: A and B are each independently for each occurrence O, N(R N ) or S; X and Y are each independently for each occurrence H, OH, a hydroxyl protecting group, a phosphate group, a phosphodiester group, an activated phosphate group, an activated phosphite group, a phosphoramidite, a solid support, —P(Z′)(Z″)O-nucleoside, —P(Z′)(Z″)O-oligonucleotide, a lipid, a PEG, a steroid, a lipophile, a polymer, —P(Z′)(Z″)O-Linker-OP(Z′″)(Z″)O-oligonucleotide, a nucleotide, an oligonucleotide, —P(Z′)(Z″)-formula (I), —P(Z′)(Z″)- or -Linker-R; R is L G , -Linker-L G , or has the structure shown below: L G is independently for each occurrence a carbohydrate, e.g., monosaccharide, disaccharide, trisaccharide, tetrasaccharide, oligosaccharide, polysaccharide; R N is independently for each occurrence H, methyl, ethyl, propyl, isopropyl, butyl, or benzyl; and Z′, Z″, Z′″ and Z″″ are each independently for each occurrence O or S.
1. A modified HBV-targeting oligonucleotide comprising a structure shown below:
wherein L G is independently for each occurrence a ligand, and all of the nucleotide sugars of the oligonucleotide are modified; and
wherein the oligonucleotide targets the HBV gene.
2. A modified HBV-targeting oligonucleotide comprising a structure shown in formula (II)-(V):
wherein:
q 2A , q 2B , q 3A , q 3B , q 4A , q 4B , q 5A , q 5B and q 5C represent independently occurrence 0-20 and wherein the repeating unit can be the same or different;
P 2A , P 2B , P 3A , P 3B , P 4A , P 4B , P 5A , P 5B , P 5C , T 2A , T 2B , T 3A , T 3B , T 4A , T 4B , T 4A , T 5B , and T 5C are each independently for each occurrence absent, CO, NH, O, S, OC(O), NHC(O), CH 2 , CH 2 NH or CH 2 O;
Q 2A , Q 2B , Q 3A , Q 3B , Q 4A , Q 4B , Q 5A , Q 5B , and Q 5C are independently for each occurrence absent, alkylene, substituted alkylene wherein one or more methylenes can be interrupted or terminated by one or more of O, S, S(O), SO 2 , N(R N ), C(R′)═C(R″), C≡C or C(O);
R 2A , R 2B , R 3A , R 3B , R 4A , R 4B , R 5A , R 5B , and R 5C are each independently for each occurrence absent, NH, O, S, CH 2 , C(O)O, C(O)NH, NHCH(R a )C(O), —C(O)—CH(R a )—NH—, CO, CH═N—O,
or heterocyclyl;
L 2A , L 2B , L 3A , L 3B , L 4A , L 4B , L 5A , L 5B and L 5C are each independently for each occurrence a carbohydrate;
R′ and R″ are each independently H, C 1 -C 6 alkyl, OH, SH or N(R N ) 2 ;
R N is independently for each occurrence methyl, ethyl, propyl, isopropyl, butyl or benzyl; and
R a is H or amino acid side chain,
wherein all of the nucleotide sugars of the oligonucleotide are modified; and
wherein the oligonucleotide targets the HBV gene.
3. The modified oligonucleotide of claim 1 , wherein the modified oligonucleotide is double stranded.
4. The modified oligonucleotide of claim 3 , wherein the structure is at the 3′-end of one of the strands.
5. The modified oligonucleotide of claim 4 , wherein the structure is at the 3′-end of the sense strand.
6. The modified oligonucleotide of claim 1 , wherein the structure is
7. The modified oligonucleotide agent of claim 1 , wherein the structure is
8. The modified oligonucleotide of claim 1 , wherein L G is carbohydrate.
9. The modified oligonucleotide of claim 8 , wherein the carbohydrate is selected from the group consisting of monosaccharide, disaccharide, trisaccharide, tetrasaccharide and polysaccharide.
10. The modified oligonucleotide of claim 1 , wherein L G is galactose or N-acetylgalactosamine.
11. The modified oligonucleotide of claim 1 , wherein at least one of the nucleotide sugars of the modified oligonucleotide is modified at the 2′ position to replace 2′ hydroxyl.
12. The modified oligonucleotide of claim 11 , wherein the 2′ hydroxyl is replaced with a moiety selected from the group consisting of 2′-H, 2′-methoxyethyl, 2′-OCH 3 , 2′-O-allyl, 2′-C-allyl, and 2′-fluoro.
13. The modified oligonucleotide of claim 1 , wherein the oligonucleotide comprises 2′-deoxy, 2′-methoxyethyl, and phosphorothioate linkage.
14. The modified oligonucleotide of claim 1 , wherein the oligonucleotide comprises 2′-deoxy, LNA, and phosphorothioate linkage.
15. The modified oligonucleotide of claim 1 , wherein the modified oligonucleotide is single stranded.
16. The modified oligonucleotide of claim 15 , wherein the modified oligonucleotide is a hairpin or antisense.
17. A method of modulating the expression of a target HBV gene in a cell, comprising providing to said cell the modified oligonucleotide of claim 1 .
18. A pharmaceutical composition comprising the modified oligonucleotide of claim 1 alone or in combination with a pharmaceutically acceptable carrier or excipient.
19. A modified HBV-targeting oligonucleotide comprising a structure of
wherein all of the nucleotide sugars of the oligonucleotide are modified, and wherein the oligonucleotide targets the HBV gene.
20. A modified HBV-targeting oligonucleotide comprising a structure of
wherein all of the nucleotide sugars of the oligonucleotide are modified, and wherein the oligonucleotide targets the HBV gene.