Compositions and methods for inhibition of expression of protein C (PROC) genes

Patent metadata

Field	Value
Publication number	US-9725718-B2
Application number	US-201514730100-A
Country	US
Kind code	B2
Filing date	Jun 3, 2015
Priority date	Jun 21, 2011
Publication date	Aug 8, 2017
Grant date	Aug 8, 2017

Abstract

The invention relates to double-stranded ribonucleic acid (dsRNA) targeting a PROC gene, and methods of using the dsRNA to inhibit expression of PROC.

First claim

We claim:

1. A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of a Protein C (PROC) gene, wherein the dsRNA comprises a sense strand and an antisense strand each 30 nucleotides or less in length, wherein the antisense strand is complementary to at least 15 contiguous nucleotides of nucleotides SEQ ID NO:159.

2. A double-stranded ribonucleic acid (dsRNA) for inhibiting expression of gene, wherein the dsRNA comprises a sense strand and an antisense strand wherein the sense strand consists of the nucleotide sequence of the sense strand of AD-48898.1UM (SEQ ID NO:159) and the antisense strand consists of the nucleotide sequence of the antisense strand of AD-48898.1UM (SEQ ID NO:213).

3. The dsRNA of claim 1 , wherein the antisense strand comprises the antisense strand of AD-48898.1UM (SEQ ID NO:213).

4. The dsRNA of claim 1 , wherein at least one nucleotide of the dsRNA is a modified nucleotide.

5. The dsRNA of claim 4 , wherein the modified nucleotide is chosen from the group consisting of: a 2′-O-methyl modified nucleotide, a nucleotide comprising a 5′-phosphorothioate group, and a terminal nucleotide linked to a cholesteryl derivative or dodecanoic acid bisdecylamide group.

6. The dsRNA of claim 4 , wherein the modified nucleotide is chosen from the group consisting of: a 2′-deoxy-2′-fluoro modified nucleotide, a 2′-deoxy-modified nucleotide, a locked nucleotide, an abasic nucleotide, 2′-amino-modified nucleotide, 2′-alkyl-modified nucleotide, morpholino nucleotide, a phosphoramidate, and a non-natural base comprising nucleotide.

7. The dsRNA of claim 1 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.

8. The dsRNA of claim 1 , wherein each strand comprises a 3′ overhang of at 2 nucleotides.

9. The dsRNA of claim 1 , further comprising a ligand.

10. The dsRNA of claim 9 , wherein the ligand is conjugated to the 3′ end of the sense strand of the dsRNA.

11. The dsRNA of claim 1 , further comprising at least one N-Acetyl-Galactosamine.

12. A cell comprising the dsRNA of claim 1 .

13. A vector encoding at least one strand of the dsRNA of claim 1 .

14. A cell comprising the vector of claim 13 .

15. A pharmaceutical composition for inhibiting expression of an PROC gene comprising the dsRNA of claim 1 .

16. The pharmaceutical composition of claim 15 , comprising a lipid formulation.

17. The pharmaceutical composition of claim 15 , comprising a lipid formulation comprising (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (MC3).

18. A method of inhibiting PROC expression in a cell, the method comprising:
(a) contacting the cell the dsRNA of claim 1 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of an PROC gene, thereby inhibiting expression of the PROC gene in the cell.

19. The method of claim 18 , wherein the PROC expression is inhibited by at least 30%.

20. A method of treating a disorder mediated by PROC expression comprising administering to a human in need of such treatment a therapeutically effective amount of the PROC dsRNA of claim 1 .

21. The method of claim 20 , wherein the disorder is a bleeding disorder.

22. The method of claim 20 , wherein the disorder is hemophilia.

23. The method of claim 20 , wherein administration causes an increase in blood clotting and/or a decrease in PROC protein accumulation.

24. The method of claim 20 , wherein the dsRNA or the pharmaceutical composition is administered at a dose of about 0.01 mg/kg to about 10 mg/kg or about 0.5 mg/kg to about 50 mg/kg.

25. A pharmaceutical composition for inhibiting expression of an PROC gene comprising the dsRNA of claim 2 .

26. The pharmaceutical composition of claim 25 , comprising a lipid formulation.

27. The pharmaceutical composition of claim 25 , comprising a lipid formulation comprising (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (MC3).

28. A method of inhibiting PROC expression in a cell, the method comprising:
(a) contacting the cell the dsRNA of claim 2 ; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of an PROC gene, thereby inhibiting expression of the PROC gene in the cell.

29. A method of treating a disorder mediated by PROC expression comprising administering to a human in need of such treatment a therapeutically effective amount of the PROC dsRNA of claim 2 .

30. The method of claim 29 , wherein the disorder is a bleeding disorder.

31. The method of claim 29 , wherein the disorder is hemophilia.

32. The method of claim 29 , wherein administration causes an increase in blood clotting and/or a decrease in PROC protein accumulation.

33. The method of claim 29 , wherein the dsRNA or the pharmaceutical composition is administered at a dose of about 0.01 mg/kg to about 10 mg/kg or about 0.5 mg/kg to about 50 mg/kg.

Assignees

• Alnylam Pharmaceuticals Inc

Inventors

• Toudjarska Ivanka
• Maraganore John M
• Bettencourt Brian
• Milstein Stuart
• Maier Martin A
• Charisse Klaus
• Rajeev Kallanthottathil
• Kuchimanchi Satyanarayana

Classifications

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N15/113 Primary

  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

• C12N2310/3515

  Lipophilic moiety, e.g. cholesterol · CPC title

• C12N2310/321

  2'-O-R Modification · CPC title

• C12N2310/3521

  Methyl · CPC title