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

• US 8,618,277 B2
• Filed: 05/25/2012
• Issued: 12/31/2013
• Est. Priority Date: 02/20/2002
• Status: Expired due to Term

• Current Assignee: Sirna Therapeutics, Inc. (Alnylam Pharmaceuticals Incorporated)
• Original Assignee: Merck Sharp & Dohme Corporation (Merck & Co., Inc.)
• Inventors: Beigelman, Leonid, McSwiggen, James
• Primary Examiner(s): Bowman, Amy
• Application Number:
  US13/480,655
• Publication Number:
  US 20130018082A1
• Time in Patent Office: 585 Days
• Field of Search: None
• US Class Current: 536/24.5
• 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 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/141 MicroRNAs, miRNAs
• CPC Class Codes: C12N 2310/321 2'-O-R Modification
• CPC Class Codes: C12N 2310/3521 Methyl

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 short interfering RNA (siRNA) molecule having a sense strand and an antisense strand that mediates RNA interference against a Hepatitis B Virus target RNA, wherein:

(a) each strand is between 18 and 24 nucleotides in length;

(b) the sense strand comprises 10 or more 2′

-deoxy, 2′

-O-methyl, 2′

-deoxy-2′

-fluoro, or universal base modified nucleotides;

(c) the antisense strand comprises 10 or more 2′

-deoxy, 2′

-O-methyl, 2′

-deoxy-2′

-fluoro, or universal base modified nucleotides; and

(d) 10 or more pyrimidine nucleotides of the sense and/or antisense strand are 2′

-deoxy, 2′

-O-methyl or 2′

-deoxy-2′

-fluoro nucleotides.

2. The siRNA molecule of claim 1, wherein the sense strand comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 phosphorothioate internucleotide linkages.

3. The siRNA molecule of claim 1, wherein the antisense strand comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 phosphorothioate internucleotide linkages.

4. A composition comprising the siRNA molecule of claim 1 and a pharmaceutically acceptable carrier or diluent.