Compounds and methods for enhanced cellular uptake

Patent metadata

Field	Value
Publication number	US-9506030-B2
Application number	US-201414266127-A
Country	US
Kind code	B2
Filing date	Apr 30, 2014
Priority date	May 1, 2013
Publication date	Nov 29, 2016
Grant date	Nov 29, 2016

Abstract

Described herein are conjugated modified oligonucleotides that are complementary to a target RNA. The conjugate facilitates cellular uptake of the modified oligonucleotide, resulting improved potency.

First claim

What is claimed:

1. A compound comprising the structure:

wherein each N of N m is, independently, a modified or unmodified nucleoside and m is from 1 to 5; X 1 and X 2 are each, independently, a phosphodiester linkage or a phosphorothioate linkage; and MO is a modified oligonucleotide; and wherein when m is greater than 1, each modified or unmodified nucleoside of N m is connected to adjacent modified or unmodified nucleosides of N m by a phosphodiester internucleoside linkage or a phosphorothioate internucleoside linkage.

2. The compound of claim 1 , wherein at least one of X 1 and X 2 is a phosphodiester linkage.

3. The compound of claim 1 , wherein each of X 1 and X 2 is a phosphodiester linkage.

4. The compound of claim 1 , wherein m is 1, 2, or 3.

5. The compound of claim 1 , wherein N m is N′ p N″, wherein each N′ is, independently, a modified or unmodified nucleoside and p is from 0 to 4; and N″ is a nucleoside comprising an unmodified sugar moiety; and wherein each pair of adjacent N′ nucleosides are connected by a phosphodiester internucleoside linkage or a phosphorothioate internucleoside linkage and N″ is connected to an adjacent N′ by a phosphodiester internucleoside linkage or a phosphorothioate internucleoside linkage.

6. The compound of claim 5 , wherein p is 0, 1, 2, 3, or 4.

7. The compound of claim 6 , wherein p is 1, 2, 3, or 4 and each N of N m is linked by a phosphodiester internucleoside linkage.

8. The compound of claim 5 , wherein each N′ comprises an unmodified sugar moiety.

9. The compound of claim 5 , wherein N″ is a β-D-deoxyriboadenosine or a β-D-deoxyriboguanosine.

10. The compound of claim 1 , wherein the sugar moiety of each N of N m is independently selected from a β-D-ribose, a β-D-deoxyribose, a 2′-O-methoxy sugar, a 2′-O-methyl sugar, a 2′-fluoro sugar, and a bicyclic sugar moiety.

11. The compound of claim 1 , wherein the nucleobase sequence of the modified oligonucleotide is complementary to a target RNA.

12. The compound of claim 11 , wherein the target RNA is selected from a microRNA, a messenger RNA, a pre-messenger RNA, and a long non-coding RNA.

13. The compound of claim 12 , wherein the modified oligonucleotide is hybridized to a second modified oligonucleotide, wherein the nucleobase sequence of the second modified oligonucleotide is complementary to the nucleobase sequence of the modified oligonucleotide.

14. The compound of claim 11 , wherein the target RNA is a human target RNA.

15. The compound of claim 1 , wherein the nucleobase sequence of the modified oligonucleotide is at least 90% identical to the nucleobase sequence of a microRNA.

16. The compound of claim 15 , wherein the microRNA is a human microRNA.

17. The compound of claim 1 , wherein the modified oligonucleotide consists of 8 to 25 linked nucleosides.

18. The compound of claim 1 , wherein the modified oligonucleotide comprises at least one nucleoside with a modified sugar moiety.

19. The compound of claim 18 , wherein each modified sugar moiety is independently selected from a 2′-O-methyl sugar moiety, a 2′-O-methoxyethyl sugar moiety, a 2′-fluoro sugar moiety, and a bicyclic sugar moiety.

20. The compound of claim 19 , wherein each bicyclic sugar moiety is independently selected from a cEt sugar moiety and an LNA sugar moiety.

21. The compound of claim 1 , wherein each nucleoside of the modified oligonucleotide comprises a modified sugar moiety.

22. The compound of claim 1 , wherein the modified oligonucleotide comprises a plurality of nucleosides with a modified sugar moiety, and a plurality of nucleosides with an unmodified sugar moiety.

23. The compound of claim 1 , wherein the modified oligonucleotide comprises a plurality of non-bicyclic nucleosides and a plurality of bicyclic nucleosides.

24. The compound of claim 1 , wherein at least one linkage of the modified oligonucleotide is a modified internucleoside linkage.

25. A process of making a compound comprising the structure

wherein each N of N m is, independently, a modified or unmodified nucleoside and m is from 1 to 5; X 1 and X 2 are each, independently, a phosphodiester linkage or a phosphorothioate linkage; wherein when m is greater than 1, each modified or unmodified nucleoside of N m is connected to adjacent modified or unmodified nucleosides of N m by a phosphodiester internucleoside linkage or a phosphorothioate internucleoside linkage; and MO is a modified oligonucleotide, the process comprising the steps of:
providing a solid support containing a conjugate as shown in formula IV;

deprotecting the DMTr group under conditions effective to produce a reactive hydroxyl;
performing sequential phosphoramidite coupling steps to form N m ;
performing sequential phosphoramidite coupling steps to form the modified oligonucleotide; and

releasing the conjugated modified oligonucleotide from the solid support.

Assignees

• Regulus Therapeutics Inc

Inventors

• Bhat Balkrishen

Classifications

• A61P1/16

  for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics · CPC title

• C12N2310/351

  Conjugate · CPC title

• C12N15/111

  General methods applicable to biologically active non-coding nucleic acids · CPC title

• C12N2310/31

  of the backbone · CPC title

• C12N2310/11

  Antisense · CPC title