Serpina1 iRNA compositions and methods of use thereof

Patent metadata

Field	Value
Publication number	US-11725207-B2
Application number	US-202117194431-A
Country	US
Kind code	B2
Filing date	Mar 8, 2021
Priority date	Nov 23, 2016
Publication date	Aug 15, 2023
Grant date	Aug 15, 2023

Abstract

The invention relates to RNAi agents, e.g., double stranded RNAi agents, targeting the Serpina1 gene, and methods of using such RNAi agents to inhibit expression of Serpina1 and methods of treating subjects having a Serpina1 associated disease, such as a liver disorder.

First claim

We claim:

1. A method of inhibiting serine peptidase inhibitor, Glade A, member 1 (Serpina1) expression in a cell, the method comprising contacting the cell with a double stranded RNA (dsRNA) agent comprising a sense strand and an antisense strand forming a double stranded region,
wherein the antisense strand comprises at least 19 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of

(SEQ ID NO: 419)

5′-UUUUGUUCAAUCAUUAAGAAGAC-3′,

wherein all of the nucleotides of the antisense strand comprise a nucleotide modification,
wherein the antisense strand comprises at least one thermally destabilizing modification selected from the group consisting of an abasic modification, a mismatch with the opposing nucleotide in the opposing strand, a 2′-deoxy modification, and an acyclic nucleotide modification at position 4, 5, 6, 7 and/or 8 from the 5′-end,
wherein the nucleotides at positions 2, 6, 8, 9, 14, and 16, or at positions 2, 6, 14, and 16, or at positions 2, 14, and 16 from the 5′-end of the antisense strand comprise a 2′-F nucleotide modification,
wherein all of the nucleotides of the sense strand comprise a nucleotide modification,
wherein the nucleotides at positions 7, 9, 10, and 11, or at positions 7, 10, and 11 from the 5′-end of the sense strand comprise a 2′-F nucleotide modification,
wherein the sense strand comprises an ASGPR ligand, and
wherein each of the sense strand and the antisense strand are independently 19 to 25 nucleotides in length,
thereby inhibiting Serpina1 expression in the cell.

2. A method of inhibiting serine peptidase inhibitor, Glade A, member 1 (Serpina1) expression in a cell, the method comprising contacting the cell with a double stranded RNA (dsRNA) agent comprising a sense strand and an antisense strand forming a double stranded region,
wherein the antisense strand comprises at least 19 contiguous nucleotides differing by no more than 3 nucleotides from the nucleotide sequence of

(SEQ ID NO: 419)

5′-UUUUGUUCAAUCAUUAAGAAGAC-3′,

wherein all of the nucleotides of the antisense strand comprise a nucleotide modification,
wherein the thermally destabilizing modification is located in position 4 to 8 from the 5′-end of the antisense strand and is selected from the group consisting of an abasic modification, a mismatch with the opposing nucleotide in the opposing strand, a 2′-deoxy modification, and an acyclic nucleotide modification,
wherein the nucleotides at positions 2, 6, 8, 9, 14, and 16, or at positions 2, 6, 14, and 16, or at positions 2, 14, and 16 from the 5′-end of the antisense strand comprise a 2′-F nucleotide modification,
wherein all of the nucleotides of the sense strand comprise a nucleotide modification,
wherein the nucleotides at positions 7, 9, 10, and 11, or at positions 7, 10, and 11 from the 5′-end of the sense strand comprise a 2′-F nucleotide modification,
wherein each of the sense strand and the antisense strand are independently 19 to 25 nucleotides in length, and
wherein the dsRNA agent has a melting temperature of from about 40° C. to about 80° C.,
thereby inhibiting Serpina1 expression in the cell.

3. The method of claim 1 or 2 , wherein said cell is within a subject.

4. The method of claim 1 or 2 , wherein the subject is a human.

5. The method of claim 1 or 2 , wherein the thermally destabilizing modification is selected from the group consisting of

wherein B is nucleobase.

6. The method of claim 1 or 2 , wherein the destabilizing modification is located at position 7 of the antisense strand.

7. The method of claim 1 or 2 , wherein all of the nucleotides of the sense strand and all of the nucleotides of the antisense strand comprise a nucleotide modification selected from the group consisting of a thermally destabilizing modification, a 2′-F modification and a 2′-OMe modification.

8. The method of claim 2 , wherein the dsRNA agent further comprises an ASGPR ligand.

9. The method of claim 1 or 8 , wherein the ASGPR ligand is one or more GalNAc derivatives attached through a monovalent, bivalent or trivalent linker.

10. The method of claim 1 or 8 , wherein the ASGPR ligand is:

11. The method of claim 10 , wherein the dsRNA agent is conjugated to the ligand as shown in the following schematic

wherein X is O or S.

12. The method of claim 1 or 2 , wherein each strand is independently 19 to 23 nucleotides in length.

13. The method of claim 1 or 2 , wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.

14. The method of claim 1 or 2 , wherein the nucleotides at positions 2, 14, and 16 from the 5′-end of the antisense strand comprise a 2′-F nucleotide modification.

15. The method of claim 1 or 2 , wherein the nucleotides at positions 7, 9, 10, and 11 from the 5′-end of the sense strand comprise a 2′-F modified nucleotide modification.

16. The method of claim 1 or 2 , wherein the sense strand further comprises two phosphorothioate or methylphosphonate internucleotide linkages.

17. The method of claim 1 or 2 , wherein the two phosphorothioate or methylphosphonate internucleotide linkages are located between nucleotide positions 1 and 2 and between nucleotide positions 2 and 3 from the 5′-end of the sense strand.

18. The method of claim 1 or 2 , wherein the antisense strand further comprises four phosphorothioate or methylphosphonate internucleotide linkages.

19. The method of claim 18 , wherein the four phosphorothioate or methylphosphonate internucleotide linkages are located between nucleotide positions 1 and 2, between nucleotide positions 2 and 3, between nucleotide positions 21 and 22, and between nucleotide positions 22 and 23 from the 5′-end of the antisense strand.

20. The method of claim 1 or 2 , wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide.

21. The method of claim 1 or 2 , wherein the antisense strand differs by no more than 2 nucleotides from the nucleotide sequence of

(SEQ ID NO: 419)

5′-UUUUGUUCAAUCAUUAAGAAGAC-3′.

22. The method of claim 1 or 2 , wherein the antisense strand differs by no more

Assignees

• Alnylam Pharmaceuticals Inc

Inventors

• Schlegel Mark K
• Janas Maja
• Jadhav Vasant R
• Foster Donald
• Manoharan Muthiah
• Rajeev Kallanthottathil G
• Kel'In Alexander V
• Charisse Klaus
• Nair Jayaprakash K
• Maier Martin A
• Matsuda Shigeo
• Jayaraman Muthusamy
• Sehgal Alfica
• Brown Christopher
• Fitzgerald Kevin
• Milstein Stuart

Classifications

• 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

• C12N2310/14

  interfering nucleic acids [NA] · CPC title

• C12N2310/315

  Phosphorothioates · CPC title

• C12N2310/351

  Conjugate · CPC title

• A61K48/00 Primary

  Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy · CPC title