Two-tailed self-delivering siRNA

Patent metadata

Field	Value
Publication number	US-10844377-B2
Application number	US-201816015440-A
Country	US
Kind code	B2
Filing date	Jun 22, 2018
Priority date	Jun 23, 2017
Publication date	Nov 24, 2020
Grant date	Nov 24, 2020

Abstract

Provided are compositions and methods comprising two-tailed siRNAs (tt-siRNAs) that exhibit unprecedented cellular uptake and silencing. Also provided are methods of treating neurological and other diseases with the two-tailed siRNAs of the invention.

First claim

What is claimed is:

1. A double-stranded nucleic acid compound comprising:
a) a sense strand having a 5′ end, a 3′ end and a region of complementarity with an antisense strand;
b) an antisense strand having a 5′ end, a 3′ end, a region of complementarity with target RNA;
c) a first overhang region at the 3′ end of the sense strand having at least 4 contiguous phosphorothioated nucleotides; and
d) a second overhang region at the 3′ end of the antisense strand having at least 4 contiguous phosphorothioated nucleotides,
wherein the nucleotides at positions 1 and 2 from the 5′ end of the sense and antisense strands are connected to adjacent nucleotides via phosphorothioate linkages.

2. The compound of claim 1 , wherein the antisense strand comprises a 5′ phosphate moiety.

3. The compound of claim 1 , wherein the antisense strand comprises a moiety Rat the 5′ end, wherein R is selected from the group consisting of:

4. The compound of claim 1 , wherein the sense strand and the anti sense strand each independently comprise at least 15 contiguous nucleotides.

5. The compound of claim 1 , wherein the overhang regions of the sense strand and the antisense strand independently comprise 2′-methoxy-nucleotides and 2′-fluoro-nucleotides.

6. The compound of claim 5 , wherein the overhang region of the sense strand and the antisense strand independently consists of at least four consecutive 2′-methoxy-nucleotides.

7. The compound of claim 6 , wherein the nucleotides at positions 1, 2, 3, and 4 from the 3′ end of the sense and antisense strands consist of 2′-methoxy-nucleotides.

8. The compound of claim 1 , wherein the nucleotides at positions 1-7 or 1-8 from the 3′ end of the sense strand or the 3′ end of the antisense strand, independently, are connected to adjacent nucleotides via phosphorothioate linkages.

9. The compound of claim 1 , wherein the overhang regions of the sense strand and the antisense strand have the same number of phosphorothioated nucleotides.

10. The compound of claim 3 , having the structure selected from Formulas (I-VIII):

wherein:
X, for each occurrence, independently, is selected from adenosine, guanosine, uridine, cytidine, and chemically-modified derivatives thereof;
Y, for each occurrence, independently, is selected from adenosine, guanosine, uridine, cytidine, and chemically-modified derivatives thereof;
- represents a phosphodiester internucleoside linkage;
= represents a phosphorothioate internucleoside linkage;
--- represents, individually for each occurrence, a base-pairing interaction or a mismatch; and
R, for each occurrence, is a nucleotide comprising a 5′ phosphate or is R1, R2, R3, R4, R5, R6, R7 or R8, as defined above.

11. The compound of claim 1 , wherein the antisense strand has between 80% and 99% complementarity to the target RNA.

12. A pharmaceutical composition comprising one or more double stranded nucleic acid compounds of claim 1 , and a pharmaceutically acceptable carrier.

13. A method of treating a disease or disorder comprising administering to a subject in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 12 .

14. The method of claim 13 , wherein the subject in need of such treatment is a human.

15. A method for selective in vivo delivery of a compound of claim 1 to a target organ, tissue or cells, comprising administering the compound to a subject.

16. A method of treating a neurological disease or disorder comprising administering to a subject in need of such treatment a therapeutically effective amount of the pharmaceutical composition of claim 12 .

17. A method of treating a neurological disease or disorder comprising administering to a subject in need of such treatment a therapeutically effective amount of the double stranded nucleic acid compound having the structure of Formula (I) or Formula (VI), or Formula (IV) or Formula (VII) of claim 10 .

18. The method of claim 17 , wherein the pharmaceutical composition is administered by intravenous injection, intraperitoneal injection, intracranial injection, intrathecal injection, intrastriatal injection, or intracerebroventricular injection.

19. A double-stranded nucleic acid compound comprising
a) a sense strand having a 5′ end, a 3′ end and a region of complementarity with an antisense strand;
b) an antisense strand having a 5′ end, a 3′ end, a region of complementarity with target RNA;
c) a first overhang region at the 3′ end of the sense strand having 7 contiguous phosphorothioated nucleotides; and
d) a second overhang region at the 3′ end of the antisense strand having 7 contiguous phosphorothioated nucleotides.

20. A double-stranded nucleic acid compound comprising:
a) a sense strand having a 5′ end, a 3′ end and a region of complementarity with an antisense strand;
b) an antisense strand having a 5′ end, a 3′ end, a region of complementarity with target RNA;
c) a first overhang region at the 3′ end of the sense strand comprising at least 4 contiguous phosphorothioated nucleotides; and
d) a second overhang region at the 3′ end of the antisense strand comprising at least 5 contiguous phosphorothioated nucleotides.

21. The compound of claim 5 , wherein the overhang regions of the sense strand and the antisense strand consist of 2′-methoxy-nucleotides.

22. The compound of claim 1 , wherein the overhang region of the sense strand and the antisense strand independently consists of 4, 5, 6, 7, or 8 phosphorothioated nucleotides.

23. The compound of claim 1 , wherein the overhang regions of the sense strand and the antisense strand have different numbers of phosphorothioated nucleotides.

24. The compound of claim 1 , wherein the overhang comprises abasic nucleotides.

25. The compound of claim 1 , wherein the antisense strand has perfect complementarity to the target RNA.

26. The method of claim 16 , wherein the neurological disease or disorder is Huntington's disease.

27. The method of claim 15 , wherein the target organ is the brain.

28. The method of claim 15 , wherein the target cells are primary cortical neurons.

29. The method of claim 15 , wherein the delivery of the compound is not mediated by lipid formulation.

30. The method of claim 15 , wherein the compound is administered by intravenous injection, intraperitoneal injection, intracranial injection, intrathecal injection, intrastriatal injection, or intracerebroventricular injection.

Assignees

• Univ Massachusetts

Inventors

• Khvorova Anastasia
• Alterman Julia
• Hassler Matthew

Classifications

• C12N2310/343

  having patterns, e.g. ==--==--==-- · CPC title

• C12N2310/315

  Phosphorothioates · CPC title

• C12N2310/321

  2'-O-R Modification · CPC title

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N2310/322

  2'-R Modification · CPC title