Carbohydrate conjugates as delivery agents for oligonucleotides

Patent metadata

Field	Value
Publication number	US-9352048-B2
Application number	US-201414338161-A
Country	US
Kind code	B2
Filing date	Jul 22, 2014
Priority date	Dec 4, 2007
Publication date	May 31, 2016
Grant date	May 31, 2016

Abstract

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.

First claim

We claim:

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.

Assignees

• Alnylam Pharmaceuticals Inc

Inventors

• Manoharan Muthiah
• Rajeev Kallanthottathil G
• Nair Jayaprakash
• Maier Martin

Classifications

• A61P37/02

  Immunomodulators · CPC title

• A61P3/06

  Antihyperlipidemics · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P35/00

  Antineoplastic agents · CPC title

• A61P25/00

  Drugs for disorders of the nervous system · CPC title