Carbohydrate conjugates as delivery agents for oligonucleotides

1. A phosphorothioate-modified oligonucleotide comprising a structure shown below:

wherein:

L G is independently for each occurrence a monosaccharide carbohydrate selected from the group consisting of allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, D-fucitol, L-fucitol, fucose, fucosamine, fuculose, galactose, D-galactosaminitol, galactosamine, N-acetyl-galactosamine (GalNAc), glucose, glucose-6- phosphate, glucosaminitol, glucosamine, N-acetyl-glucosamine (GluNAc), glucose glyceraldehyde, L-glycero-D-manno-heptose, glycerol, glycerone, gulose, idose, lyxose, mannose, mannose-6-phosphate, mannosamine, lactose, psicose, quinovose, quinovosamine, rhamnose, rhamnitol, rhamnosamine, ribose, ribulose, sedoheptulose, sorbose, tagatose, talose, tartaric acid, threose, xylose, and xylulose;

the Linker is —[(P-Q″-R) q -X-(P′-Q ′″-R′)] q″ -T-, wherein:

P, R, T, P R′, and T are each independently for each occurrence absent, CO, NH, O, S, OC(O), NHC(O), C(O)NH, CH 2 , CH 2 NH, CH 2 O; NHCH(R a )C(O), —C(O)—CH(R a )—NH—, CH═N—O,

or heterocyclyl;

Q″ and Q′″ are each independently for each occurrence absent, —(CH 2 ) n —, —C(R 1 )(R 2 )(CH 2 ) n— , —(CH 2 ) n C(R 1 )(R 2 )—, —(CH 2 CH 2 O) m CH 2 CH 2 —,or —(CH 2 CH 2 O) m CH 2 CH 2 NH—;

X is absent or a cleavable linking group;

R a is H or an amino acid side chain;

R 1 and R 2 are each independently for each occurrence H, CH 3 , OH, SH or N(R N ) 2 ;

R N is independently for each occurrence H, methyl, ethyl, propyl, isopropyl, butyl or benzyl;

q, q′, and q″ are each independently for each occurrence 0-20 and wherein the repeating unit can be the same or different;

n is independently for each occurrence 1-20; and

m is independently for each occurrence 0-50; and

the oligonucleotide is modified with at least one phosphorothioate linkage.

2. The phosphorothioate-modified oligonucleotide of claim 1 , wherein the oligonucleotide is double-stranded.

3. The phosphorothioate-modified oligonucleotide of claim 1 , wherein the oligonucleotide is single-stranded.

4. The phosphorothioate-modified oligonucleotide of claim 3 , wherein the oligonucleotide is a hairpin or antisense.

5. The phosphorothioate-modified oligonucleotide of claim 1 , wherein the structure is at the 5′-end of the oligonucleotide.

6. The phosphorothioate-modified oligonucleotide of claim 1 , wherein the monosaccharide carbohydrate is galactose or N—acetylgalactosamine.

7. The phosphorothioate-modified oligonucleotide of claim 1 , wherein the phosphorothioate-modified oligonucleotide contains nucleotide sugars having a moiety at the 2′-position selected from the group consisting of 2′-H, 2′-hydroxyl, 2′-methoxyethyl, 2′OCH 3 , 2′-O-allyl, 2′-C-allyl, and 2′-fluoro.

8. A phosphorothioate-modified oligonucleotide comprising a structure having formula (CIII):

wherein:

X is O, S, NR N or CR P 2 ;

B is independently for each occurrence hydrogen, optionally substituted natural or non—natural nucleobase, optionally substituted natural nucleobase conjugated with -linker-R L or optionally substituted non-natural nucleobase conjugated with -linker-R L ;

R 1 , R 2 , R 3 , R 4 and R 5 are each independently for each occurrence H, OR 6 , F, N(R N ) 2 , or-J-linker-R L ;

J is absent, 0, S, NR N , OC(O)NH, NHC(O)O, C(O)NH, NHC(O), NHSO, NHSO 2 , NHSO 2 NH, OC(O), C(O)O, OC(O)O, NHC(O)NH, NHC(S)NH, OC(S)NH, OP(N(R P ) 2 )O, or OP(N(R P ) 2 );

R 6 is independently for each occurrence hydrogen, hydroxyl protecting group, optionally substituted alkyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted aralkyl, optionally substituted alkenyl, optionally substituted heteroaryl, polyethyleneglycol (PEG), a phosphate, a diphosphate, a triphosphate, a phosphonate, a phosphonothioate, a phosphonodithioate, a phosphorothioate, a phosphorothiolate, a phosphorodithioate, a phosphorothiolothionate, a phosphodiester, a phosphotriester, an activated phosphate group, an activated phosphite group, a phosphoramidite, a solid support, —P(Z 1 )(Z 2 )—O-nucleoside, —P(Z 1 )(Z 2 )—O-oligonucleotide, —P(Z 1 )(Z 2 )-formula (CIII), —P(Z 1 )(O-linker-R L )—O-nucleoside, —P(Z 1 )(O-linker-R L )—O-oligonucleotide, or —P(Z 1 )(O-linker-R L )-O-formula (CIII);

R N is independently for each occurrence H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted aralkyl, optionally substituted heteroaryl or an amino protecting group;

R P is independently for each occurrence occurrence H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted cycloalkyl or optionally substituted heteroaryl;

R L is hydrogen or a ligand; and

Z 1 and Z 2 are each independently for each occurrence O, S N(alkyl) or optionally substituted alkyl;

provided that i) R 6 is a phosphorothioate at least once; and ii) R 5 is-J-linker-R L at least once, wherein R L is a ligand having the structure of

wherein L G is independently for each occurrence a monosaccharide carbohydrate selected from the group consisting of allose, altrose, arabinose, cladinose, erythrose, erythrulose, fructose, D-fucitol, L-fucitol, fucose, fucosamine, fuculose, galactose, D-galactosaminitol, galactosamine, N-acetyl-galactosamine (GalNAc), glucose, glucose-6-phosphate, glucosaminitol, glucosamine, N-acetyl-glucosamine (GluNAc), glucose glyceraldehyde, L-glycero-D-manno-heptose, glycerol, glycerone, gulose, idose, lyxose, mannose, mannose-6-phosphate, mannosamine, lactose, psicose, quinovose, quinovosamine, rhamnose, rhamnitol, rhamnosamine, ribose, ribulose, sedoheptulose, sorbose, tagatose, talose, tartaric acid, threose, xylose, and xylulose;

wherein the linker in formula (CIII) and the Linker in the structure of R L each independently is —[(P-Q″-R) q -X-(P′-Q′″-R′) q′ ] q″ -T-, wherein:

P, R, T, P′, R′, and T are each independently for each occurrence absent, CO, NH, O, S, OC(O), NHC(O), C(O)NH, CH 2 , CH 2 NH, CH 2 O; NHCH(R a )C(O), —C(O)—CH(R a )—NH—, CH═N-O,

or heterocyclyl;

Q″ and Q′″ are each independently for each occurrence absent, —(CH 2 ) n —, —C(R 1 )(R 2 )(CH 2 ) n —, —(CH 2 C(R 1 )(R 2 )—, —(CH 2 CH 2 O) m CH 2 CH 2 —, or —(CH 2 CH 2 O) m CH 2 CH 2 NH—;

X is absent or a cleavable linking group;

R a is H or an amino acid side chain;

R 1 and R 2 are each independently for each occurrence H, CH 3 , OH, SH or N(R N ) 2 ;

R N is independently for each occurrence H, methyl, ethyl, propyl, isopropyl, butyl or benzyl;

q, q′, and q″ are each independently for each occurrence 0-20 and wherein the repeating unit can be the same or different;

n is independently for each occurrence 1-20; and

m is independently for each occurrence 0-50.

9. The phosphorothioate-modified oligonucleotide of claim 1 , wherein the oligonucleotide is double-stranded.

10. The phosphorothioate-modified oligonucleotide of claim 8 , wherein the oligonucleotide is single-stranded.

11. The phosphorothioate-modified oligonucleotide of claim 10 , wherein the oligonucleotide is a hairpin or antisense.

12. The phosphorothioate-modified oligonucleotide of claim 8 , wherein the monosaccharide carbohydrate is galactose or N-acetylgalactosamine.