Gapped oligomeric compounds comprising 5′-modified deoxyribonucleosides in the gap and uses thereof

Patent metadata

Field	Value
Publication number	US-9752142-B2
Application number	US-201214238094-A
Country	US
Kind code	B2
Filing date	Aug 8, 2012
Priority date	Aug 11, 2011
Publication date	Sep 5, 2017
Grant date	Sep 5, 2017

Abstract

The present invention provides gapped oligomeric compounds comprising at least one 5′-substituted P-D-2′-deoxyribonucleoside in the gap region. Certain such gapped oligomeric compounds are useful for hybridizing to a complementary nucleic acid, including but not limited to, nucleic acids in a cell. The oligomeric compounds provided herein have improved properties such as selectivity, potency and improved proinflammatory profile. In certain embodiments, hybridization results in modulation of the amount of activity or expression of the target nucleic acid in a cell.

First claim

The invention claimed is:

1. A gapped oligomeric compound comprising a contiguous sequence of linked monomer subunits having a gap region located between a 5′-region and a 3′-region wherein the 5′ and 3′-regions each, independently, have from 2 to 8 contiguous RNA-like
modified nucleosides that each adopt a 3′-endo conformational geometry when put into an oligomeric compound and wherein the gap region has from 6 to 14 contiguous monomer subunits selected from β-D-2′-deoxyribonucleosides and 5′-substituted β-D-2′-deoxyribonucleosides having Formula I:

wherein independently for each 5′-substituted β-D-2′-deoxyribonucleoside having Formula I:
T 1 and T 2 are each, independently, an internucleoside linking group linking the 5′-substituted β-D-2′-deoxyribonucleoside having Formula I to the remainder of the gapped oligomeric compound;
Bx is a heterocyclic base moiety;
R 1 is C 1 -C 6 alkyl, substituted C 1 -C 6 alkyl, C 2 -C 6 alkenyl, substituted C 2 -C 6 alkenyl, C 2 -C 6 alkynyl or substituted C 2 -C 6 alkynyl; and
wherein said gap region comprises at least one 5′-substituted β-D-2′-deoxyribonucleoside having Formula I and a plurality of β-D-2′-deoxyribonucleosides.

2. The gapped oligomeric compound of claim 1 wherein each substituted group comprises one or more optionally protected substituent groups independently selected from halogen, OJ 1 , NJ 1 J 2 , ═NJ 1 , SJ 1 , N 3 , CN, OC(=L)J 1 , OC(=L)NJ 1 J 2 and NJ 3 C(=L)NJ 1 J 2 , wherein each J 1 , J 2 and J 3 is, independently, H, C 1 -C 6 alkyl or a protecting group, and L is O, S or NJ 1 .

3. The gapped oligomeric compound of claim 1 wherein each R 1 is independently selected from C 1 -C 6 alkyl, substituted C 1 -C 6 alkyl, C 2 -C 6 alkenyl and C 2 -C 6 alkynyl.

4. The gapped oligomeric compound of claim 1 wherein each R 1 is independently selected from CH 3 , CH 2 CH 3 , CH 2 CH 2 OH, CH 2 CHCH 2 and CHCH 2 .

5. The gapped oligomeric compound of claim 1 wherein each R 1 is CH 3 .

6. The gapped oligomeric compound of claim 1 having only one 5′-substituted β-D-2′-deoxyribonucleoside of Formula I.

7. The gapped oligomeric compound of claim 1 having only two 5′-substituted β-D-2′-deoxyribonucleosides of Formula I.

8. The gapped oligomeric compound of claim 1 wherein each 5′-substituted β-D-2′-deoxyribonucleoside has the configuration of Formula Ia:

9. The gapped oligomeric compound of claim 1 wherein each 5′-substituted β-D-2′-deoxyribonucleoside has the configuration of Formula Ib:

10. The gapped oligomeric compound of claim 1 wherein each internucleoside linking group that links adjacent monomer subunits is independently selected from a phosphodiester or phosphorothioate internucleoside linking group.

11. The gapped oligomeric compound of claim 1 wherein each internucleoside linking group that links adjacent monomer subunits is a phosphorothioate internucleoside linking group.

12. The gapped oligomeric compound of claim 1 wherein each monomer subunit comprises a heterocyclic base moiety independently selected from uracil, thymine, cytosine, 4-N-benzoylcytosine, 5-methylcytosine, 4-N-benzoyl-5-methylcytosine, adenine, 6-N-benzoyladenine, guanine and 2-N-isobutyrylguanine.

13. The gapped oligomeric compound of claim 1 wherein each modified nucleoside in the 5′ and 3′-regions is independently selected from a bicyclic nucleoside comprising a bicyclic furanosyl sugar moiety or a modified nucleoside comprising a furanosyl sugar moiety having at least one substituent group.

14. The gapped oligomeric compound of claim 13 comprising one or more 2′-modified nucleosides that each have a 2′-substituent group independently selected from halogen, OCH 3 , OCH 2 F, OCHF 2 , OCF 3 , OCH 2 CH 3 , O(CH 2 ) 2 F, OCH 2 CHF 2 , OCH 2 CF 3 , OCH 2 —CH═CH 2 , O(CH 2 ) 2 —OCH 3 , O(CH 2 ) 2 —SCH 3 , O(CH 2 ) 2 —OCF 3 , O(CH 2 ) 3 —N(R 3 )(R 4 ), O(CH 2 ) 2 —ON(R 3 )(R 4 ), O(CH 2 ) 2 —O(CH 2 ) 2 —N(R 3 )(R 4 ), OCH 2 C(═O)—N(R 4 )(R 4 ), OCH 2 C(═O)—N(R 5 )—(CH 2 ) 2 —N(R 3 )(R 4 ) and O(CH 2 ) 2 —N(R 5 )—C(═NR 6 )[N(R 3 )(R 4 )] wherein R 3 , R 4 , R 5 and R 6 are each, independently, H or C 1 -C 6 alkyl.

15. The gapped oligomeric compound of claim 14 wherein each 2′-substituent group is independently selected from F, OCH 3 , O(CH 2 ) 2 —OCH 3 and OCH 2 C(═O)—N(H)CH 3 .

16. The gapped oligomeric compound of claim 15 wherein each 2′-substituent group is O(CH 2 ) 2 —OCH 3 .

17. The gapped oligomeric compound of claim 13 comprising one or more bicyclic nucleosides wherein each bicyclic nucleoside has a bridging group independently selected from 4′-(CH 2 )—O-2′, 4′-(CH 2 )—S-2′, 4′-(CH 2 ) 2 —O-2′, 4′-CH(CH 3 )—O-2′, 4′-CH(CH 2 OCH 3 )—O-2′, 4′-C(CH 3 ) 2 —O-2′, 4′-CH 2 —N(OCH 3 )-2′, 4′-CH 2 —O—N(CH 3 )-2′, 4′-CH 2 —NCH 3 —O-2′, 4′-CH 2 —C(H)(CH 3 )-2′ and 4′-CH 2 —C(═CH 2 )-2′.

18. The gapped oligomeric compound of claim 17 wherein each bridging group is 4′-CH[(S)—(CH 3 )]—O-2′.

19. The gapped oligomeric compound of claim 1 comprising at least two different types of modified nucleosides in the 5′ and 3′-regions comprising at least two different modified ribofuranosyl sugar moieties.

20. The gapped oligomeric compound of claim 19 wherein the 5′ and 3′-regions include only 4′-CH[(S)—(CH 3 )]—O-2′ bicyclic nucleosides and 2′-O(CH 2 ) 2 —OCH 3 substituted nucleosides.

21. The gapped oligomeric compound of claim 1 wherein the modified nucleosides in the 5′ and 3′-regions each have the same modified sugar moiety.

22. The gapped oligomeric compound of claim 1 wherein the 5′ and 3′-regions each, independently, have from 3 to 6 monomer subunits and the gap region has from 6 to 10 monomer subunits.

23. A method of inhibiting gene expression comprising contacting one or more cells, a tissue or the animal with the oligomeric compound of claim 1 wherein said oligomeric compound is complementary to a target RNA.

24. The method of claim 23 wherein said cells are in a human.

25. The method of claim 23 wherein said target RNA is human mRNA.

26. The method of claim 23 wherein said target RNA is cleaved thereby inhibiting the function of said target RNA.

Assignees

• Oestergaard Michael
• Prakash Thazha P
• Seth Punit P
• Swayze Eric E
• Ionis Pharmaceuticals Inc

Inventors

• Oestergaard Michael
• Prakash Thazha P
• Seth Punit P
• Swayze Eric E

Classifications

• C12N15/111

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

• C12N2310/312

  Phosphonates · CPC title

• C12N2320/34

  Allele or polymorphism specific uses · CPC title

• C12N2310/3341

  5-Methylcytosine · CPC title

• C12N2310/11

  Antisense · CPC title