Tetragalnac and peptide containing conjugates and methods for delivery of oligonucleotides

1. A modular composition comprising:

1) a single stranded or double stranded oligonucleotide;

2) one or more tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different:

wherein X is —O—, —S—, —CR 1 R 2 — or —NR 1 —, wherein R 1 and R 2 are each independently selected from the group consisting of hydrogen and C1-C6 alkyl; n is 1, 2, 3, or 4; and the bond with “ ” indicates the point of attachment;

optionally, 3) one or more linkers, which may be the same or different;

optionally, 4) one or more peptides independently selected from SEQ ID No. 1-474, or the D-amino acid, retro-inverso, and cysteine conjugation point variants thereof, which may be the same or different, wherein the cysteine conjugation point variants thereof refers to variants of the peptides comprising conjugation through existing cysteines or through a cysteine residue added to a N- or C-terminus of the peptides; and

optionally, 5) one or more targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.

2. The modular composition of claim 1 comprising:

1) a single stranded or double stranded oligonucleotide;

2) 1-8 tetraGalNAc ligands of Formula (II), which may be the same or different, wherein X is —O—, —S—, —CH 2 — or NH—; and n is 1, 2, 3, or 4;

3) 1-24 linkers, which may be the same or different;

4) 1-8 peptides independently selected from SEQ ID No. 1-474, which may be the same or different; and

optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.

3. A modular composition comprising:

1) a single stranded or double stranded siRNA;

2) 1-8 tetraGalNAc ligands of Formula (I), (II) or (III), which may be the same or different, wherein X is —O—, —S—, —CH 2 — or NH—; and n is 1, 2, 3, or 4;

3) 1-24 linkers, which may be the same or different;

4) 1-12 peptides independently selected from SEQ ID No. 1-474, or the D-amino acid, retro-inverso, and cysteine conjugation point variants thereof, which may be the same or different, wherein the cysteine conjugation point variants thereof refers to variants of the peptides comprising conjugation through existing cysteines or through a cysteine residue added to a N- or C-terminus of the peptides; and

optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.

4. The modular composition of claim 3 , wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA optionally via linkers.

5. The modular composition of claim 3 , wherein X of Formula (I), (II) or (III) is —O—, —S— or —CH 2 —; and n is 1, 2 or 3.

6. The modular composition of claim 3 , wherein the composition comprises 1-4 tetraGalNAc ligands, which may be the same or different.

7. The modular composition of claim 3 , wherein the composition comprises 1-8 peptides, which may be the same or different.

8. The modular composition of claim 3 ,

wherein the siRNA is double stranded; and

wherein the tetraGalNAc ligands are attached to the guide strand or the passenger strand of the siRNA at different 2′-positions of the ribose rings of the siRNA.

9. The modular composition of claim 3 ,

wherein the siRNA is double stranded; and

wherein the tetraGalNAc ligands are attached to the guide strand or the passenger strand of the siRNA at different terminal 3′ and/or 5′-positions.

10. The modular composition of claim 3 ,

wherein the siRNA is double stranded; and

wherein the tetraGalNAc ligands are attached to both the guide strand and the passenger strand of the siRNA at different 2′-positions of the ribose rings and/or different terminal 3′ and/or 5′-positions.

11. The modular composition of claim 3 ,

wherein the siRNA is double stranded; and

wherein the peptides are attached to the guide strand or the passenger strand of the siRNA at different 2′-positions of the ribose rings of the siRNA.

12. The modular composition of claim 3 ,

wherein the siRNA is double stranded; and

wherein the peptides are attached to the guide strand or the passenger strand of the siRNA at different terminal 3′ and/or 5′-positions.

13. The modular composition of claim 3 ,

wherein the siRNA is double stranded; and

wherein the peptides are attached to both the guide strand and the passenger strand of the siRNA at different 2′-positions of the ribose rings and/or different terminal 3′ and/or 5′-positions.

14. The modular composition of claim 3 , wherein the tetraGalNAc ligands and the peptides are attached to the same strand of the siRNA.

15. The modular composition of claim 3 , wherein the tetraGalNAc ligands and the peptides are attached to different strands of the siRNA.

16. The modular composition of claim 3 , wherein the tetraGalNAc ligands and the peptides are attached to the same or different strands of the siRNA via linkers.

17. The modular composition of claim 16 , wherein each linker is independently selected from the group consisting of:

wherein:

each R is independently H, Boc (tert-butyloxycarbonyl), Cbz (carboxybenzyl), Ac (acetyl), a PEG, a lipid, a targeting ligand, linker(s), or peptide(s); and

each n is 0 to 750.

18. The modular composition of claim 16 , wherein each linker is independently selected from the group consisting of:

wherein:

each R is independently H, Boc (tert-butyloxycarbonyl), Cbz (carboxybenzyl), Ac (acetyl), a PEG, a lipid, a targeting ligand, linker(s), or peptide(s); and

each n is 0 to 750.

19. The modular composition of claim 18 , wherein the linker is a branching linker independently selected from the group consisting of:

wherein:

each R is independently H, Boc (tert-butyloxycarbonyl), Cbz (carboxybenzyl), Ac (acetyl), a PEG, a lipid, a targeting ligand, linker(s), or peptide(s); and

each n is 0 to 750.

20. The modular composition of claim 3 ,

wherein the siRNA is double stranded; and

wherein the optional targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents are attached to the same or different strands of the siRNA via linkers.

21. A modular composition comprising:

1) a double stranded siRNA;

2) 1-8 tetraGalNAc ligands of Formula (IV), (V) or (VI):

3) 1-24 linkers which may be the same or different;

4) 1-12 peptides independently selected from SEQ ID No. 1-474, or the D-amino acid, retro-inverso, and cysteine conjugation point variants thereof, which may be the same or different, wherein the cysteine conjugation point variants thereof refers to variants of the peptides comprising conjugation through existing cysteines or through a cysteine residue added to a N- or C-terminus of the peptides; and,

optionally, 5) 1-8 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.

22. The modular composition of claim 21 comprising:

1) a double stranded siRNA;

2) 1-4 tetraGalNAc ligands of Formula (V);

3) 1-12 linkers which may be the same or different;

4) 1-8 peptides independently selected from SEQ ID No. 1-474, or the D-amino acid, retro-inverso, and cysteine conjugation point variants thereof, which may be the same or different, wherein the cysteine conjugation point variants thereof refers to variants of the peptides comprising conjugation through existing cysteines or through a cysteine residue added to a N- or C-terminus of the peptides; and,

optionally, 5) 1-4 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents;

wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and

wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA optionally via linkers.

23. The modular composition of claim 21 , wherein the tetraGalNAc ligands and the peptides are attached to the same strand of the siRNA via linkers.

24. The modular composition of claim 21 , wherein the tetraGalNAc ligands and the peptides are attached to different strands of the siRNA via linkers.

25. The modular composition of claim 21 comprising:

1) a double stranded siRNA;

2) 1-4 tetraGalNAc ligands of Formula (IV);

3) 1-12 linkers which may be the same or different;

4) 1-8 peptides independently selected from SEQ ID No. 2, 3, 5, 7, 11, 13, 19, 22, 27-32, 55, 56, 63, 64, 69, 71-74, 86, 90, 94, 95, 106, 137, 192, 200, 201, 228, 229, 266, 282, 333, 337, 407, 423, 436, 437, 461-463, 467, 468, 470, 473 and 474, or the D-amino acid, retro-inverso, and cysteine conjugation point variants thereof, which may be the same or different, wherein the cysteine conjugation point variants thereof refers to variants of the peptides comprising conjugation through existing cysteines or through a cysteine residue added to a N- or C-terminus of the peptides; and,

optionally, 5) 1-4 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents;

wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and

wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA via linkers.

26. The modular composition of claim 25 comprising the D-amino acids of the peptides independently selected from SEQ ID No. 2, 3, 5, 7, 11, 13, 19, 22, 27-32, 55, 56, 63, 64, 69, 71-74, 86, 90, 94, 95, 106, 137, 192, 200, 201, 228, 229, 266, 282, 333, 337, 407, 423, 436, 437, 461-463, 467, 468, 470, 473 and 474, which may be the same or different.

27. The modular composition of claim 25 , wherein the tetraGalNAc ligands and the peptides are attached to the same or different strands of the siRNA via linkers.

28. The modular composition of claim 25 , wherein the tetraGalNAc ligands and the peptides are attached to different strands of the siRNA via linkers.

29. The modular composition of claim 21 comprising:

1) a double stranded siRNA;

2) 1-4 tetraGalNAc ligands of Formula (V);

3) 1-12 linkers which may be the same or different;

4) 1-8 peptides independently selected from SEQ ID No. 2, 3, 5, 7, 11, 13, 19, 22, 27-32, 55, 56, 63, 64, 69, 71-74, 86, 90, 94, 95, 106, 137, 192, 200, 201, 228, 229, 266, 282, 333, 337, 407, 423, 436, 437, 461-463, 467, 468, 470, 473 and 474, or the D-amino acid, retro-inverso, and cysteine conjugation point variants thereof, which may be the same or different, wherein the cysteine conjugation point variants thereof refers to variants of the peptides comprising conjugation through existing cysteines or through a cysteine residue added to a N- or C-terminus of the peptides; and,

optionally, 5) 1-4 targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents;

wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA at different 2′-positions of the ribose rings and/or at different terminal 3′ and/or 5′-positions of the siRNA; and

wherein the tetraGalNAc ligands and/or the peptides are attached to the siRNA via linkers.

30. The modular composition of claim 29 , wherein the tetraGalNAc ligands and the peptides are attached to the same or different strands of the siRNA via linkers.

31. The modular composition of claim 29 comprising 1 tetraGalNAc ligand, wherein the tetraGalNAc ligand is attached to the siRNA via a linker.

32. The modular composition of claim 29 comprising 1 peptide, wherein the peptide is attached to the siRNA via a linker.

33. The modular composition of claim 29 comprising 2-4 tetraGalNAc ligands, wherein the tetraGalNAc ligands are attached to the same strand or different strands of the siRNA via linkers.

34. The modular composition of claim 29 comprising 2-4 peptides, wherein the peptides are attached to the same strand or different strands of the siRNA via linkers.

35. A pharmaceutical composition comprising the modular composition of claim 1 and a pharmaceutically acceptable excipient.