RNA interference mediated inhibition of gene expression using chemically modified short interfering nucleic acid (siNA)

• US 20060276635A1
• Filed: 08/04/2006
• Published: 12/07/2006
• Est. Priority Date: 09/05/2002
• Status: Active Grant

• Current Assignee: Sirna Therapeutics, Inc. (Alnylam Pharmaceuticals Incorporated)
• Original Assignee: Sirna Therapeutics, Inc. (Alnylam Pharmaceuticals Incorporated)
• Inventors: MacEjak, Dennis, Thompson, James, Beigelman, Leonid, Chen, Tongqian, Wang, Weimin, McSwiggen, James, Vargeese, Chandra, Zinnen, Shawn, Haeberli, Peter, Morrissey, David, Usman, Nassim, Jamison, Sharon, Chowrira, Bharat, Fosnaugh, Kathy, Vaish, Narendra, Pavco, Pamela
• Granted Patent: US 7,956,176 B2
• Time in Patent Office: Days
• US Class Current: 536/23.100
• CPC Class Codes: A61K 47/544 Phospholipids
• CPC Class Codes: A61K 47/549 Sugars, nucleosides, nucleotides or nucleic acids
• CPC Class Codes: A61K 47/554 the modifying agent being a steroid plant sterol, glycyrrhetic acid, enoxolone or bile acid
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/0048 Eye, e.g. artificial tears
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 31/16 for influenza or rhinoviruses
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 31/22 for herpes viruses
• CPC Class Codes: C12N 15/111 General methods applicable to biologically active non-coding nucleic acids
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/317 with an inverted bond, e.g. a cap structure
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/322 2'-R Modification
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/351 Conjugate
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2320/51 : modulating the chemical stability, e.g. nuclease-resistance

Abstract

The present invention concerns methods and reagents useful in modulating gene expression in a variety of applications, including use in therapeutic, diagnostic, target validation, and genomic discovery applications. Specifically, the invention relates to synthetic chemically modified small nucleic acid molecules, such as short interfering nucleic acid (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), micro-RNA (miRNA), and short hairpin RNA (shRNA) molecules capable of mediating RNA interference (RNAi) against target nucleic acid sequences. The small nucleic acid molecules are useful in the treatment of any disease or condition that responds to modulation of gene expression or activity in a cell, tissue, or organism.

Claims

1. A chemically modified double stranded short interfering nucleic acid (siNA) molecule that mediates RNA interference (RNAi) against a mammalian RNA sequence, wherein:

a. the siNA comprises a sense strand and an antisense strand;

b. each strand of the siNA molecule is about 18 to about 27 nucleotides in length;

c. the antisense strand comprises a nucleotide sequence of about 18 to about 27 nucleotides that is complementary to the mammalian RNA sequence;

d. the sense strand comprises a portion of about 18 to about 27 nucleotides of the mammalian RNA sequence; and

e. 100% of the nucleotide positions of the sense strand, the antisense strand, or both the sense strand and antisense strand comprise chemically modified nucleotides.

2. The siNA molecule of claim 1, wherein one or more of the chemically modified nucleotides comprise a 2′

-O-methyl nucleotide.

3. The siNA molecule of claim 1, wherein one or more of the chemically modified nucleotides comprise a 2′

-deoxy-2′

-fluoro nucleotide.

4. The siNA molecule of claim 1, wherein one or more of the chemically modified nucleotides comprise a 2′

-deoxy nucleotide.

5. The siNA molecule of claim 1, wherein one or more of the chemically modified nucleotides comprise an abasic moiety.

6. The siNA molecule of claim 1, wherein one or more of the chemically modified nucleotides comprise a phosphorothioate linked nucleotide.

7. The siNA molecule of claim 1, wherein the chemically modified nucleotides comprise a combination of 2′

-O-methyl and 2′

-deoxy-2′

-fluoro nucleotides.

8. The siNA molecule of claim 1, wherein the chemically modified nucleotides comprise a combination of 2′

-O-methyl, 2′

-deoxy-2′

-fluoro, and 2′

-deoxy nucleotides.

9. The siNA molecule of claim 1, wherein the chemically modified nucleotides comprise a combination of 2′

-O-methyl, 2′

-deoxy-2′

-fluoro, 2′

-deoxy, abasic, and phosphorothioate linked nucleotides.

10. The siNA molecule of claim 1, wherein the chemically modified nucleotides comprise a combination of 2′

-O-methyl, 2′

-deoxy-2′

-fluoro, 2′

-deoxy, and phosphorothioate linked nucleotides.