NZ767118B2 - Modified double-stranded rna agents - Google Patents
Modified double-stranded rna agents Download PDFInfo
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- NZ767118B2 NZ767118B2 NZ767118A NZ76711815A NZ767118B2 NZ 767118 B2 NZ767118 B2 NZ 767118B2 NZ 767118 A NZ767118 A NZ 767118A NZ 76711815 A NZ76711815 A NZ 76711815A NZ 767118 B2 NZ767118 B2 NZ 767118B2
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- dsrna agent
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
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- A—HUMAN NECESSITIES
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- C12N15/09—Recombinant DNA-technology
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- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—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
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- C12N2310/32—Chemical structure of the sugar
- C12N2310/323—Chemical structure of the sugar modified ring structure
- C12N2310/3231—Chemical structure of the sugar modified ring structure having an additional ring, e.g. LNA, ENA
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- C12N2310/00—Structure or type of the nucleic acid
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- C12N2310/00—Structure or type of the nucleic acid
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- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/34—Spatial arrangement of the modifications
- C12N2310/344—Position-specific modifications, e.g. on every purine, at the 3'-end
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- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/34—Spatial arrangement of the modifications
- C12N2310/346—Spatial arrangement of the modifications having a combination of backbone and sugar modifications
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- C12N2310/00—Structure or type of the nucleic acid
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- C12N2310/35—Nature of the modification
- C12N2310/351—Conjugate
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- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/50—Physical structure
- C12N2310/53—Physical structure partially self-complementary or closed
- C12N2310/533—Physical structure partially self-complementary or closed having a mismatch or nick in at least one of the strands
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- C12N2320/50—Methods for regulating/modulating their activity
- C12N2320/52—Methods for regulating/modulating their activity modulating the physical stability, e.g. GC-content
Abstract
One aspect of the present invention relates to double-stranded RNA (dsRNA) agent of Formula (Is) capable of inhibiting the expression of a target gene. The sense strand of the dsRNA agent comprises at least one thermally destabilizing nucleotide, and at least one said thermally destabilizing nucleotide occurring at a site opposite to the seed region (positions 2-8) of the antisense strand, comprising four 2’-F modifications at positions 7 and 9-11 from the 5’-end of the sense strand. Other aspects of the invention relates to pharmaceutical compositions comprising these dsRNA agents suitable for therapeutic use, and methods of inhibiting the expression of a target gene by administering these dsRNA agents, e.g., for the treatment of various disease conditions.
Claims (29)
1. A double-stranded RNA (dsRNA) agent capable of inhibiting the expression of a target gene, comprising a sense strand sequence and an antisense strand sequence complementary to at least one portion of a mRNA of the target gene, each of the sense and antisense strands having 19- 5 25 nucleotides in length, wherein the sense strand is represented by formula (Is): wherein: B1, B2, and B3 each independently represent a nucleotide ning a modification selected from the group consisting of 2’-Oalkyl, 2’-substituted , 2’-substituted alkyl, 2’-halo, ENA, and 10 BNA/LNA; C1 is a thermally destabilizing nucleotide, selected from the group consisting of i) a nucleotide that forms a ch pair with the ng tide in the antisense strand, ii) a nucleotide having an abasic modification, and iii) a nucleotide having a sugar modification, and placed at a site opposite to the seed region (positions 2-8) of the antisense strand; 15 T1 represents a nucleotide sing a 2’-F modification; n1 or n3 is ndently 4 to 15 nucleotides in length; n5 is 1-6 nucleotide(s) in length; n2 is 3 nucleotides in length; n4 is 0-3 nucleotide(s) in length; and 20 wherein the sense strand has 2’-F modifications, and the 2’-F modifications on the sense strand consist of four, and only four, 2’-F modifications, at positions 7 and 9-11 from the 5’-end of the sense strand.
2. The dsRNA agent of claim 1, wherein n4 is 1.
3. The dsRNA agent of claim 2, wherein C1 is at position 14-17 of the 5’-end of the sense strand, when the sense strand is 19-22 nucleotides in .
4. The dsRNA agent of any one of claims 1 to 3, wherein C1 has thermally destabilizing 5 modification selected from the group consisting of abasic modification selected from the group consisting of: ; and sugar modification selected from the group consisting of: , , , , , , and 10 , wherein B is a modified or unmodified base, R1 and R2 independently are H, halogen, OR3, or alkyl; and R3 is H, alkyl, cycloalkyl, aryl, l, heteroaryl or sugar.
5. The dsRNA agent of any one of claims 1 to 4, wherein B1, B2, and B3 each contain 2’- OMe modifications.
6. The dsRNA agent of any one of claims 1 to 5, wherein the sense strand comprises one block of two phosphorothioate or methylphosphonate internucleotide linkages.
7. The dsRNA agent of any one of claims 1 to 6, wherein the dsRNA agent has a 3’ and/or 5’ overhang(s) of 1-10 nucleotides in length. 5
8. The dsRNA agent of any one of claims 1 to 7, further comprising at least one ASGPR ligand attached to the 3’ end of the sense .
9. The dsRNA agent of claim 8, wherein the ASGPR ligand is one or more GalNAc derivatives attached h a bivalent or trivalent branched linker.
10. The dsRNA agent of claim 9, wherein the ASGPR ligand is: HO OH O H H HO O N N O HO OH O H H HO O N N O O O O HO OH HO O N N O AcHN H H O .
11. The dsRNA agent of any one of claims 1 to 10, wherein formula (Is) further comprises a 15 2’-deoxythymidine linked via a phosphorodithioate (PS2) linkage or a yl phosphonate (VP) at the 5’-end of the sense or antisense strand.
12. The dsRNA agent of claim 1, wherein n4 is 0. 20
13. The dsRNA agent of claim 1, wherein each B1, B2, and B3 is independently 2’-OMe or 2’-F.
14. The dsRNA agent of claim 1, wherein C1 is at position 14-17 of the 5’-end of the sense strand, n4 is 1, and C1 is a glycol c acid (GNA).
15. The dsRNA agent of claim 1, wherein C1 is at position 14-17 of the 5’-end of the 5 sense strand, n4 is 1, C1 is a glycol nucleic acid (GNA), and each B1, B2, and B3 is independently 2’-OMe or 2’-F.
16. The dsRNA agent of claim 1, wherein n4 is 0, and each B1, B2, and B3 is independently 2’-OMe or 2’-F.
17. The dsRNA agent of claim 1, wherein each B1 is independently 2’-OMe or 2’-F, and each B2 and B3 is 2’-OMe.
18. The dsRNA agent of claim 1, wherein each B2 and B3 is 2’-OMe.
19. A pharmaceutical composition comprising the dsRNA agent according to any one of claims 1 to 18 in combination with a pharmaceutically acceptable r or excipient.
20. Use of the dsRNA agent according to any one of claims 1 to 18 in the manufacture of a 20 medicament for inhibiting the expression of a target gene in a human subject.
21. The use of claim 20, wherein the dsRNA agent is prepared for subcutaneous or enous administration.
22. A method for inhibiting the expression of a target gene in vitro or in a non-human subject, comprising the step of administering the dsRNA agent according to any one of claims 1 to 18 in an amount sufficient to inhibit expression of the target gene. 5
23. The method of claim 22, n the dsRNA agent is administered through subcutaneous or intravenous stration.
24. Use of the dsRNA agent according to any one of claims 1 to 18 in the manufacture of a medicament for delivering a cleotide to a ic target in a human t.
25. The use of claim 24, wherein said dsRNA agent is prepared for administration by an administration means comprising intramuscular, intrabronchial, intrapleural, eritoneal, intraarterial, lymphatic, intravenous, subcutaneous, cerebrospinal, or combinations thereof. 15
26. A method for delivering a polynucleotide to a specific target in vitro or in a non-human subject by administering the dsRNA agent according to any one of claims 1 to 18.
27. The method of claim 26, wherein said administering is carried out by an administration means comprising intramuscular, intrabronchial, intrapleural, intraperitoneal, intraarterial, 20 lymphatic, intravenous, subcutaneous, cerebrospinal, or combinations thereof.
28. Use of the dsRNA agent according to any one of claims 1 to 18 in the cture of a medicament for delivering a polynucleotide to a specific target of a human subject, wherein the dsRNA agent is prepared for subcutaneous administration, such that the polynucleotide is 25 delivered to the specific target of the subject.
29. A method for delivering a polynucleotide to a specific target in a non-human subject, the method comprising: delivering a dsRNA agent according to any one of claims 1 to 18 by subcutaneous administration into the t, such that the polynucleotide is delivered into specific target of the subject. None set by KJM MigrationNone set by KJM Unmarked set by KJM None set by KJM MigrationNone set by KJM Unmarked set by KJM .... m mM $01."th m . 9....\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\\
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201462039507P | 2014-08-20 | 2014-08-20 | |
| US201462083744P | 2014-11-24 | 2014-11-24 | |
| US201462093919P | 2014-12-18 | 2014-12-18 | |
| NZ730296A NZ730296A (en) | 2014-08-20 | 2015-08-14 | Modified double-stranded rna agents |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| NZ767118A NZ767118A (en) | 2024-02-23 |
| NZ767118B2 true NZ767118B2 (en) | 2024-05-24 |
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