Modified short interfering nucleic acid (siNA) molecules and uses thereof

Patent metadata

Field	Value
Publication number	US-11549110-B2
Application number	US-202217672268-A
Country	US
Kind code	B2
Filing date	Feb 15, 2022
Priority date	Mar 6, 2020
Publication date	Jan 10, 2023
Grant date	Jan 10, 2023

Abstract

Disclosed herein are short interfering nucleic acid (siNA) molecules comprising modified nucleotides and uses thereof. The siNA molecules may be double stranded and comprise modified nucleotides selected from 2′-O-methyl nucleotides and 2′-fluoro nucleotides. Further disclosed herein are siNA molecules comprising (a) a phosphorylation blocker, conjugated moiety, or 5′-stabilized end cap; and (b) a short interfering nucleic acid (siNA).

First claim

What is claimed is:

1. A double stranded short interfering nucleic acid (siNA) comprising:
(a) a sense strand comprising 19-21 nucleotides in a nucleic acid sequence that is at least 80% identical to SEQ ID NO: 40, wherein 15 or more of the nucleotides are modified nucleotides independently selected from a 2′-O-methyl nucleotide and a 2′-fluoro nucleotide, and wherein at least 11 of the modified nucleotides are a 2′-O-methyl nucleotide and at least 4 of the modified nucleotides are a 2′-fluoro nucleotide; and
(b) an antisense strand comprising 19-21 nucleotides in a nucleic acid sequence that is at least 80% complementary to SEQ ID NO: 40, wherein 15 or more of the nucleotides are modified nucleotides independently selected from a 2′-O-methyl nucleotide and a 2′-fluoro nucleotide, and wherein at least 11 of the modified nucleotides are a 2′-O-methyl nucleotide and 4 to 6 of the modified nucleotides are a 2′-fluoro nucleotide.

2. The double stranded siNA of claim 1 , wherein:
(a) the nucleotide(s) at position 3, 5, 7, 8, 9, 10, 11, 12, 14, 17, or 19 from the 5′ end of the sense strand is a 2′-fluoro nucleotide; and
(b) the nucleotide(s) at position 2, 5, 6, 8, 10, 14, 16, 17, or 18 from the 5′ end of the antisense strand is a 2′-fluoro nucleotide.

3. The double stranded siNA of claim 1 , wherein the sense strand comprises SEQ ID NO: 438 or SEQ ID NO: 435, and the antisense strand comprises any one of SEQ ID NOs: 501-519, SEQ ID NO: 537, SEQ ID NO: 538, or SEQ ID NO: 539.

4. A double stranded short interfering nucleic acid (siNA) comprising:
(a) a sense strand comprising 19 nucleotides, wherein 2′-fluoro nucleotides are at positions 5 and 7-9 from the 5′ end of the sense strand, and wherein 2′-O-methyl nucleotides are at positions 1-4, 6, and 10-19 from the 5′ end of the sense strand; and
(b) an antisense strand comprising 21 nucleotides, wherein 2′-fluoro nucleotides are at positions 2, 6, 14, and 16 from the 5′ end of the antisense strand, and wherein 2′-O-methyl nucleotides are at positions 1, 3-5, 7-13, 15, and 17-21 from the 5′ end of the antisense strand.

5. The double stranded siNA of claim 4 further comprising (i) the nucleotides at positions 1 and 2 and positions 2 and 3 from the 5′ end of the sense strand are connected by phosphorothioate internucleoside linkages; and (ii) the nucleotides at positions 1 and 2; positions 2 and 3; positions 19 and 20; and positions 20 and 21 from the 5′ end of the antisense strand are connected by phosphorothioate internucleoside linkages.

6. The double stranded siNA of claim 4 further comprising a conjugated moiety attached to the sense strand.

7. The double stranded siNA of claim 6 , wherein the conjugated moiety is a galactosamine.

8. The double stranded siNA of claim 6 , wherein the conjugated moiety is N-acetylgalactosamine (GalNAc).

9. The double stranded siNA of claim 4 , wherein the sense strand comprises SEQ ID NO: 438, and the antisense strand comprises any one of SEQ ID NO: 501, SEQ ID NO: 537, SEQ ID NO: 538, or SEQ ID NO: 539.

10. The double stranded siNA of claim 4 , wherein the sense strand is at least 80% identical to SEQ ID NO: 40 and the antisense strand is at least 80% complementary to SEQ ID NO: 40.

11. The double stranded siNA of claim 4 , wherein the sense strand or the antisense strand comprises at least one overhang consisting of 1 or 2 nucleotides.

12. The double stranded siNA of claim 4 , wherein both the sense strand and the antisense strand each independently comprise an overhang consisting of 1 or 2 nucleotides, wherein the overhang on the sense strand and the overhang on the antisense strand are on opposite ends of the double stranded siNA.

13. A short interfering nucleic acid (siNA) molecule comprising:
(a) a sense strand comprising a nucleic acid sequence consisting of SEQ ID NO: 438 or SEQ ID NO: 435, and
(b) an antisense strand comprising a nucleic acid sequence consisting of any one of SEQ ID NO: 501, SEQ ID NO: 505, SEQ ID NO: 506, SEQ ID NO: 537, SEQ ID NO: 538, and SEQ ID NO: 539.

14. The double stranded siNA of claim 9 , wherein the senses strand comprises SEQ ID NO: 438 and the antisense comprises SEQ ID NO: 501.

15. The double stranded siNA of claim 9 , wherein the senses strand comprises SEQ ID NO: 438 and the antisense comprises any one of SEQ ID NO: 537-539.

16. A double stranded short interfering nucleic acid (siNA) comprising:

(a) a sense strand of

(SEQ ID NO: 438)

5′-mGpsmUpsmGmGfUmGfGfAfCmUmUmCmUmCmUmCmAmAmU-p-

ps2-GalNAc-3′

and an antisense strand of

(SEQ ID NO: 501)

5′-mApsfUpsmUmGmAfGmAmGmAmAmGmUmCfCmAfCmCmAmCpsm

GpsmA-3′;

(b) a sense strand of

(SEQ ID NO: 438)

5′-mGpsmUpsmGmGfUmGfGfAfCmUmUmCmUmCmUmCmAmAmU-p-

ps2-GalNAc-3′

and an antisense strand of

(SEQ ID NO: 521)

5′-mApsfUpsmUmGmAfGmAfGfAmAmGmUmCfCmAfCmCmAmCpsm

GpsmA-3′;

(c) a sense strand of

(SEQ ID NO: 438)

5′-mGpsmUpsmGmGfUmGfGfAfCmUmUmCmUmCmUmCmAmAmU-p-

ps2-GalNAc-3′

and an antisense strand of

(SEQ ID NO: 522)

5′-mApsfUpsmUmGfAmGmAfGmAmAmGmUmCfCmAmCfCmAmCpsm

GpsmA-3′;

(d) a sense strand of

(SEQ ID NO: 438)

5′-mGpsmUpsmGmGfUmGfGfAfCmUmUmCmUmCmUmCmAmAmU-

p-ps2-GalNAc-3′

and an antisense strand of

(SEQ ID NO: 537)

5′-mApsf4PpsmUmGmAfGmAmGmAmAmGmUmCfCmAfCmCmAmCp

smGpsmA-3′;

(e) a sense strand of

(SEQ ID NO: 438)

5′-mGpsmUpsmGmGfUmGfGfAfCmUmUmCmUmCmUmCmAmAmU-

p-ps2-GalNAc-3′

and an antisense strand of

(SEQ ID NO: 538)

5′-mApsfUpsmUmGmAfGmAmGmAmAmGmUmCf2PmAfCmCmA

Assignees

• Aligos Therapeutics Inc

Inventors

• Beigelman Leonid
• Rajwanshi Vivek Kumar
• Hossbach Markus
• Pandey Rajendra K
• Hong Jin
• Eltepu Laxman
• Montero Saul Martinez
• Costa N Tilani S De

Classifications

• C12N2310/322

  2'-R Modification · CPC title

• C12N15/1131 Primary

  against viruses · CPC title

• C12N2310/317

  with an inverted bond, e.g. a cap structure · CPC title

• C12N2310/351

  Conjugate · CPC title

• C12N2310/315

  Phosphorothioates · CPC title