Method and medicament for inhibiting the expression of a given gene

• US 8,101,584 B2
• Filed: 03/06/2003
• Issued: 01/24/2012
• Est. Priority Date: 01/30/1999
• Status: Expired due to Term

• Current Assignee: Alnylam Pharmaceuticals Incorporated
• Original Assignee: Alnylam Pharmaceuticals Incorporated
• Inventors: Kreutzer, Roland, Limmer, Stefan
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US10/382,395
• Publication Number:
  US 20040072779A1
• Time in Patent Office: 3,246 Days
• Field of Search: 514/44, 536/24.5
• US Class Current: 514/44
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• 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 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/53 partially self-complementary or closed
• CPC Class Codes: C12N 2330/30 chemically synthesised
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention provides methods and compositions for inhibiting gene expression using double stranded RNA molecules that are between 15 and 21 nucleotides in length and are complementary to a target gene sequence.

Claims

1. A method for inhibiting the expression of a target gene in a mammalian cell in vitro, comprising the steps of:

a) introducing into said cell in vitro an isolated double stranded ribonucleotide molecule (dsRNA), and

b) maintaining the cell produced in step a) for a time sufficient to obtain degradation of an RNA transcript of said target gene, thereby inhibiting the expression of said target gene in said cell, and

wherein said dsRNA molecule

i) consists of two linked RNA strands, a first RNA strand that is complementary to a mammalian target gene and a second RNA strand that is complementary to said first strand,

ii) said dsRNA consists of 21 base pairs.

2. The method of claim 1, wherein said two linked RNA strands are fully complementary to each other.

3. The method of claim 1, wherein one strand of the dsRNA is fully complementary to an RNA transcript of at least part of said target gene.

4. The method of claim 3, wherein said two linked complementary strands are fully complementary to each other.

5. The method of claim 1, wherein said RNA transcript of said target gene is a primary or processed RNA transcript.

6. The method of claim 1, wherein at least one of said strands comprises at least one chemically modified nucleotide.

7. The method of claim 1, wherein said two linked RNA strands are stabilized by a chemical linkage comprising a labile linker.

8. The method of claim 7, wherein said labile linker comprises a disulfide.

9. The method of claim 1, wherein said two linked RNA strands are stabilized by a chemical linkage comprising a covalent linker.

10. The method of claim 9, wherein said linker is a C18 linker.