Granted Patent B2

US 12,297,430 · App. 17/580,269 · Granted May 13, 2025

Inventors: Anastasia Khvorova (Westborough, MA); Julia Alterman (Worcester, MA); Sarah Davis (Boston, MA); Anton Turanov (Boston, MA)

Assignee: UNIVERSITY OF MASSACHUSETTS

C12N15/113C12N2310/14C12N2310/315C12N2310/321C12N2310/322C12N2310/52

Abstract

Claims (31)

1. An oligonucleotide comprising,

at least 14 contiguous nucleotides, a 5′ end and a 3′ end; and

at least 85% 2′-O-methyl nucleotide modifications,

wherein the nucleotides at positions 2, 14, and 20 from the 5′ end of the oligonucleotide comprise a non-2′-O-methyl modification.

2. The oligonucleotide of claim 1 , wherein the nucleotides at positions 2, 14, and 20 from the 5′ end of the oligonucleotide comprise a 2′-F modification, a 2′-H modification, or a 2′-OH moiety.

3. The oligonucleotide of claim 1 , further comprising a complementary second oligonucleotide comprising at least 13 contiguous nucleotides, and a 5′ end and a 3′ end.

4. The oligonucleotide of claim 3 , wherein the second oligonucleotide comprises between 15 and 20 contiguous nucleotides, 15 contiguous nucleotides, 18 contiguous nucleotides, or 20 contiguous nucleotides.

5. The oligonucleotide of claim 3 , wherein the second oligonucleotide comprises one or more nucleotide mismatches between the first oligonucleotide and the second oligonucleotide.

6. The oligonucleotide of claim 1 , comprising between 15 and 22 contiguous nucleotides, 20 contiguous nucleotides, or 22 contiguous nucleotides.

7. A pharmaceutical composition comprising one or more oligonucleotides of claim 1 , and a pharmaceutically acceptable carrier.

8. A method of treating or managing a disease or disorder comprising administering to a subject in need of such treatment or management a therapeutically effective amount of the pharmaceutical composition of claim 7 .

9. A double-stranded nucleic acid structure comprising an antisense strand and a sense strand, wherein:

the antisense strand comprises at least 14 contiguous nucleotides, a 5′ end and a 3′ end, and has complementarity to a target;

the sense strand comprises at least 13 contiguous nucleotides, a 5′ end and a 3′ end, and has complementarity to the antisense strand;

the antisense strand comprises at least 85% 2′-O-methyl modifications; and

the nucleotides at positions 2, 14, and 20 from the 5′ end of the antisense strand comprise a non-2′-O-methyl modification or are modified with 2′-F.

10. The double-stranded nucleic acid of claim 9 , wherein the nucleotides at positions 2, 14, and 20 from the 5′ end of the antisense strand comprise a 2′-F modification, a 2′-H modification, or a 2′-OH moiety.

11. The double-stranded nucleic acid of claim 9 , wherein the antisense strand comprises between 15 and 22 contiguous nucleotides, 20 contiguous nucleotides, or 22 contiguous nucleotides.

12. The double-stranded nucleic acid of claim 9 , wherein the sense strand comprises between 15 and 20 contiguous nucleotides, 15 contiguous nucleotides, 18 contiguous nucleotides, or 20 contiguous nucleotides.

13. The double-stranded nucleic acid of claim 9 , wherein the sense strand comprises one or more nucleotide mismatches between the antisense strand and the sense strand.

14. The double-stranded nucleic acid of claim 9 , wherein the antisense strand comprises a 5′ phosphate, a 5′-alkyl phosphonate, a 5′ alkylene phosphonate, or a 5′ alkenyl phosphonate.

15. A pharmaceutical composition comprising one or more double-stranded, chemically-modified nucleic acids of claim 9 , and a pharmaceutically acceptable carrier.

16. A method of treating or managing a disease or disorder comprising administering to a subject in need of such treatment or management a therapeutically effective amount of the pharmaceutical composition of claim 15 .

17. A branched oligonucleotide compound capable of mediating RNA silencing in a cell, comprising two or more double-stranded nucleic acids,

wherein the nucleic acids (N) are connected to one another by one or more moieties selected from a linker (L), a spacer (S), and optionally a branching point (B),

wherein each double-stranded nucleic acid comprises an antisense strand and a sense strand, wherein each antisense strand comprises at least 14 contiguous nucleotides, a 5′ end and a 3′ end, and at least one antisense strand comprises at least 85% 2′-O-methyl modifications,

wherein the nucleotides at positions 2, 14, and 20 from the 5′ end of the at least one antisense strand comprise a non-2′-O-methyl modification, and

wherein one or more nucleotides at positions 1-7 from the 3′ end of at least one antisense strand are connected to adjacent nucleotides via phosphorothioate linkages.

18. The branched oligonucleotide compound of claim 17 , wherein the compound further comprises a hydrophobic moiety attached to the terminal 5′ position of the branched oligonucleotide compound.

19. The branched oligonucleotide compound of claim 17 , wherein the non-2′-O-methyl modification comprises a 2′-F modification, a 2′-H modification, or a 2′-OH moiety.

20. The branched oligonucleotide compound of claim 17 , wherein the antisense strand comprises at least 90% 2′-O-methyl modified nucleotides.

Assignments (3)

Continuity (3)

References Cited (400)

Cited By (2)

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