Short interfering nucleic acid (siNA) molecules containing a 2′ internucleoside linkage

• US 9,896,688 B2
• Filed: 02/20/2014
• Issued: 02/20/2018
• Est. Priority Date: 02/22/2013
• Status: Active Grant

• Current Assignee: Sirna Therapeutics, Inc. (Alnylam Pharmaceuticals Incorporated)
• Original Assignee: Sirna Therapeutics, Inc. (Alnylam Pharmaceuticals Incorporated)
• Inventors: Chang, Wonsuk, Guidry, Erin N., Stanton, Matthew G., Zewge, Daniel
• Primary Examiner(s): Chong, Kimberly
• Application Number:
  US14/769,361
• Publication Number:
  US 20160068837A1
• Time in Patent Office: 1,461 Days
• Field of Search: None
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/04 specific for metastasis
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 21/02 with ribosyl as saccharide radical
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 15/113 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 when used in plants C12N15/8218
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/319 linked by 2'-5' linkages, i.e. having a free 3'-position

Abstract

The present invention relates to RNAi molecules, and compositions thereof, comprising a 2′ internucleoside linkage connecting the nucleotide at position 1 and the nucleotide at position 2 at the 5′ end of the antisense strand. Specifically, the invention relates to single- and double-stranded short interfering nucleic acid (siNA) molecules that are capable of mediating RNA interference comprising 5′ modified nucleotides that comprise, among other potential modifications, a 2′ internucleoside linkage. The invention further relates to 5′ modified nucleotides used as reagents to generate the RNAi molecules of the invention and methods of using the disclosed RNAi molecules.

Claims

1. A short interfering nucleic acid (siNA) molecule capable of mediating RNA interference comprising an antisense strand that is complementary to a nucleic acid target, wherein said antisense strand comprises a 5′

modified nucleotide having Formula II;

2. The siNA molecule of claim 1, wherein R³ is independently selected from H, hydroxy, F, Cl, C_1-6 alkyl, C_2-6 alkenyl, C_2-6 alkynyl, aryl,

3. The siNA molecule of claim 1, wherein A is —

OCH₂—

, —

CH₂CH₂—

or —

CH═

CH—

.

4. The siNA molecule of claim 1, wherein D¹ and D^1′

is independently selected from hydroxyl, —

OCH₃ or —

OCH₂CH₃.

5. The siNA molecule of claim 1, wherein B is uracil, thymine, cytosine, 5-methylcytosine, adenine or guanine.

6. The siNA molecule of claim 1, wherein E is O.

7. The siNA molecule of claim 1, wherein J is a phosphodiester internucleoside linking group or a phosphorothioate internucleoside linking group.

8. The siNA molecule of claim 1, wherein R¹ is H or hydroxyl, and R1′

;

is H, hydroxyl, halogen or —

OR⁶.

9. The siNA molecule of claim 1, wherein R¹, R^1′

and R² are each H.

10. The siNA molecule of claim 1, wherein the 5′

modified nucleotide is

11. The siNA molecule claim 10, wherein B is thymine.

12. The siNA molecule of claim 1, wherein the siNA molecule is a single-stranded, antisense molecule.

13. The siNA molecule of claim 12, wherein the siNA molecule comprises one or more additional chemically-modified nucleotides.

14. The siNA molecule of claim 1, wherein the siNA molecule is a double-stranded molecule comprising a sense strand that is complementary to the antisense strand.

15. The siNA molecule of claim 14, wherein the siNA molecule comprises one or more additional chemically-modified nucleotides.

16. A composition comprising the siNA molecule of claim 1 in a pharmaceutically acceptable carrier or diluent.