Tetragalnac and peptide containing conjugates and methods for delivery of oligonucleotides
Disclosed herein is a modular composition comprising 1) an oligonucleotide; 2) one or more tetraGalNAc ligands of Formula (I), which may be the same or different; optionally, 3) one or more linkers, which may be the same or different; 4) one or more peptides independently selected from Table 3, which may be the same or different; and optionally, 5) one or more targeting ligands, solubilizing agents, pharmacokinetics enhancing agents, lipids, and/or masking agents.
1. A method for preparing a tetraGalNAc compound having a formula:
wherein:
X is —O—, —S—, or —CH 2 —; and
n, m, and q are each 1, 2, 3, or 4,
the method comprising:
reacting an azide source having the formula of
with a compound of formula
or a compound of formula
to obtain an intermediate compound;
reacting the intermediate compound with a Lys-alkyne compound of formula
to form an acetate-protected tetraGalNAc compound; and
removing the acetate protecting groups on the acetate-protected tetraGalNAc compound, thereby forming the tetraGalNAc compound.
2. The method of claim 1 , wherein the tetraGalNAc compound has the structure of
3. The method of claim 2 , wherein m is 2 and q is 1.
4. A method for preparing a tetraGalNAc compound of formula (I):
wherein:
X is —O—, —S—, or —CH 2 —; and
n is 1, 2, 3, or 4,
the method comprising:
reacting an azide source having the formula of
with a compound of formula
or a compound of formula
to obtain an intermediate compound;
reacting the intermediate compound with a Lys-alkyne compound of formula
to form an acetate-protected tetraGalNAc compound; and
removing the acetate protecting groups on the acetate-protected tetraGalNAc compound, thereby forming the tetraGalNAc compound.
5. The method of claim 4 , wherein the tetraGalNAc compound has the structure of
wherein the Lys-alkyne compound has the formula of
6. The method of claim 5 , wherein X is —O— or —CH 2 —; and n is 1, 2, or 3.
7. The method of claim 1 , wherein the reaction to obtain the intermediate compound is carried out by reacting the azide source with a compound of formula
in the presence of trifluoromethanesulfonic acid.
8. The method of claim 1 , wherein the reaction to obtain the intermediate compound is carried out by reacting the azide source with a compound of formula
in the presence of trimethylsilyl trifluoromethanesulfonate.
9. The method of claim 1 , wherein the reaction of the intermediate compound with the Lys-alkyne compound is carried out in the presence of CuBrSMe 2 .
10. The method of claim 9 , wherein the reaction of the intermediate compound with the Lys-alkyne compound is carried out in the presence of CuBrSMe 2 and diazabicycloundecene (DBU).
11. The method of claim 1 , wherein about 4 to 6 moles of the intermediate compound are added to react with each mole of the Lys-alkyne compound.
12. The method of claim 11 , wherein about 5 to 6 moles of the intermediate compound are added to react with each mole of the Lys-alkyne compound.
13. The method of claim 1 , wherein the removing the acetate protecting groups on the acetate-protected tetraGalNAc compound is carried out by reacting the acetate-protected tetraGalNAc compound with sodium methoxide.
14. The method of claim 4 , wherein the tetraGalNAc compound has the structure of
wherein the azide source is
and the Lys-alkyne compound has the formula of
15. The method of claim 14 , wherein the tetraGalNAc compound is prepared by:
reacting
with a compound of formula
in the presence of trimethylsilyl trifluoromethanesulfonate, to obtain an intermediate compound;
reacting the intermediate compound with
in the presence of CuBrSMe 2 , to form an acetate-protected tetraGalNAc compound; and
reacting the acetate-protected tetraGalNAc compound with sodium methoxide to remove the acetate protecting groups on the acetate-protected tetraGalNAc compound, thereby forming the tetraGalNAc compound.
16. The method of claim 4 , wherein the tetraGalNAc compound has the structure of
wherein the azide source is
and the Lys-alkyne compound has the formula of
17. The method of claim 14 , wherein the tetraGalNAc compound is prepared by:
reacting
with a compound of formula
in the presence of trifluoromethanesulfonic acid, to obtain an intermediate compound of formula;
reacting the intermediate compound with
in the presence of CuBrSMe 2 , to form a acetate-protected tetraGalNAc compound; and
reacting the acetate-protected tetraGalNAc compound with sodium methoxide to remove the acetate protecting groups on the acetate-protected tetraGalNAc compound, thereby forming the tetraGalNAc compound.
18. The method of claim 4 , wherein the tetraGalNAc compound has the structure of
wherein the azide source is
and the Lys-alkyne compound has the formula of
19. The method of claim 18 , wherein the tetraGalNAc compound is prepared by:
reacting
with a compound of formula
in the presence of trifluoromethanesulfonic acid, to obtain an intermediate compound;
reacting the intermediate compound with
in the presence of CuBrSMe 2 , to form an acetate-protected tetraGalNAc compound; and
reacting the acetate-protected tetraGalNAc compound with sodium methoxide to remove the acetate protecting groups on the acetate-protected tetraGalNAc compound, thereby forming the tetraGalNAc compound.
20. The method of claim 4 , wherein the tetraGalNAc compound has the structure of
wherein the azide source is
and the Lys-alkyne compound has the formula of
21. The method of claim 20 , wherein the tetraGalNAc compound is prepared by:
reacting
with a compound of formula
in the presence of trifluoromethanesulfonic acid, to obtain an intermediate compound;
reacting the intermediate compound with
in the presence of CuBrSMe 2 , to form an acetate-protected tetraGalNAc compound; and
reacting the acetate-protected tetraGalNAc compound with sodium methoxide to remove the acetate protecting groups on the acetate-protected tetraGalNAc compound, thereby forming the tetraGalNAc compound.