Nucleic acid molecules with novel chemical compositions capable of modulating gene expression

• US 6,673,611 B2
• Filed: 05/25/2001
• Issued: 01/06/2004
• Est. Priority Date: 04/20/1998
• Status: Expired due to Term

• Current Assignee: Ribozyme Pharmaceuticals, Inc. (Alnylam Pharmaceuticals Incorporated)
• Original Assignee: Sirna Therapeutics, Inc. (Alnylam Pharmaceuticals Incorporated)
• Inventors: Thompson, James, Beigelman, Leonid, Reynolds, Mark, McSwiggen, James, Haeberli, Peter, Zwick, Michael, Matulic-Adamic, Jasenka, Jarvis, Thale, Karpeisky, Alexander, Woolf, Tod, Bellon, Laurent
• Primary Examiner(s): Shukla, Ram R.
• Assistant Examiner(s): ZARA, JANE J
• Application Number:
  US09/866,316
• Publication Number:
  US 20020142980A1
• Time in Patent Office: 956 Days
• Field of Search: 435/6, 435/91.31, 435/91.1, 435/325, 435/366, 435/375, 435/455, 536/23.1, 536/24.31, 536/24.33, 536/24.5, 514/44
• US Class Current: 435/455
• 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 35/00 Antineoplastic agents
• CPC Class Codes: A61P 5/30 Oestrogens
• 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 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 15/1138 against receptors or cell surface proteins
• CPC Class Codes: C12N 2310/15 Nucleic acids forming more than 2 strands, e.g. TFOs
• CPC Class Codes: C12N 2310/31 of the backbone
• 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 2310/332 Abasic residue
• CPC Class Codes: C12N 2310/3341 5-Methylcytosine
• CPC Class Codes: C12N 2310/341 Gapmers, i.e. of the type ===---===
• CPC Class Codes: C12N 2310/345 having at least two different backbone modifications
• CPC Class Codes: C12N 2310/346 having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/3521 Methyl
• CPC Class Codes: C12N 2310/3527 Other alkyl chain

Abstract

This invention relates to novel chemically-modified nucleic acid molecules having specific formulae that exhibit increased resistance to nucleases and increased binding affinity to target nucleic acid molecules. The invention further relates to methods of modulating gene expression using the novel chemically modified nucleic acid molecules, and compositions and cells comprising said molecules.

Claims

1. A nucleic acid molecule having the formula XIII:

2. The nucleic acid molecule of claim 1, wherein said 2′

-O-alkyl nucleotide is a 2′

-O-methyl nucleotide.

3. The nucleic acid molecule of claim 1, wherein said 2′

-O-alkyl nucleotide is a 2′

-O-allyl nucleotide.

4. The nucleic acid molecule of claim 1, wherein said inverted abasic moiety is a 3′

,3′

-linked or 5′

,5′

-linked deoxyabasic moiety.

5. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule is an antisense nucleic acid molecule.

6. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule further comprises a 2-5A antisense chimera.

7. A method of modulating the expression of a gene in a cell in culture comprising the step of administering to said cell a nucleic acid molecule of claim 1 under conditions suitable for the down regulation of said gene.

8. A mammalian cell in culture including a nucleic acid molecule of claim 1.

9. The mammalian cell of claim 8, wherein said mammalian cell is a human cell.

10. A composition comprising the nucleic acid molecule of claim 1 and a pharmaceutically acceptable carrier.

11. A method of administering to a cell in culture a nucleic acid molecule of claim 1 comprising contacting said cell with the nucleic acid molecule under conditions suitable for said administration.

12. The method of claim 11, wherein said cell is a mammalian cell.

13. The method of claim 11, wherein said cell is a human cell.

14. The method of claim 11, wherein said administration is in the presence of a delivery reagent.

15. The method of claim 14, wherein said delivery reagent is a lipid.

16. The method of claim 15, wherein said lipid is a cationic lipid.

17. The method of claim 14, wherein said delivery reagent is a liposome.