Tetragalnac and peptide containing conjugates and methods for delivery of oligonucleotides

Patent metadata

Field	Value
Publication number	US-9840531-B2
Application number	US-201715481942-A
Country	US
Kind code	B2
Filing date	Apr 7, 2017
Priority date	May 2, 2012
Publication date	Dec 12, 2017
Grant date	Dec 12, 2017

Abstract

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.

First claim

What is claimed is:

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

Assignees

• Sirna Therapeutics Inc

Inventors

• Tellers David
• Colletti Steven L
• Dudkin Vadim
• Aaronson Jeffrey
• Momose Aaron
• Tucker Thomas Joseph
• Yuan Yu
• Calati Kathleen B
• Tian Lu
• Parmar Rubina G
• Shaw Anthony W
• Wang Weimin
• Storr Rachel Anne
• Busuek Marina
• Kowtoniuk Robert A

Classifications

• A61K31/7056

  containing five-membered rings with nitrogen as a ring hetero atom · CPC title

• A61K31/713

  Double-stranded nucleic acids or oligonucleotides · CPC title

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• C12N15/113

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• A61K47/549

  Sugars, nucleosides, nucleotides or nucleic acids · CPC title