Double-stranded oligonucleotides

• US 8,815,821 B2
• Filed: 10/20/2011
• Issued: 08/26/2014
• Est. Priority Date: 02/01/2002
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Woolf, Tod
• Primary Examiner(s): WHITEMAN, BRIAN A
• Application Number:
  US13/277,957
• Publication Number:
  US 20130045520A1
• Time in Patent Office: 1,041 Days
• Field of Search: 514/44.A
• US Class Current: 514/44.A
• CPC Class Codes: A61K 31/713 Double-stranded nucleic acids or oligonucleotides
• CPC Class Codes: A61P 1/04 for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• 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 15/1135 against oncogenes or tumor suppressor genes
• CPC Class Codes: C12N 15/1137 against enzymes viral enzymes C12N15/1131; receptors C12N15/1138
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2310/321 : 2'-O-R Modification
• CPC Class Codes: C12N 2310/33 : of the base
• CPC Class Codes: C12N 2310/351 : Conjugate
• CPC Class Codes: C12N 2310/53 : partially self-complementary or closed
• CPC Class Codes: C12N 2320/31 : Combination therapy
• CPC Class Codes: C12N 2320/50 : Methods for regulating/modulating their activity
• CPC Class Codes: C12N 2320/51 : modulating the chemical stability, e.g. nuclease-resistance
• CPC Class Codes: C12Y 207/11022 : Cyclin-dependent kinase (2.7.11.22)
• CPC Class Codes: C12Y 301/03048 : Protein-tyrosine-phosphatase (3.1.3.48)

Abstract

Antisense sequences, including duplex RNAi compositions, which possess improved properties over those taught in the prior art are disclosed. The invention provides optimized antisense oligomer compositions and method for making and using the both in in vitro systems and therapeutically. The invention also provides methods of making and using the improved antisense oligomer compositions.

Claims

1. A method of decreasing expression of a target gene in a cell, comprising:

contacting the cell with a composition comprising a double-stranded oligonucleotide, the double-stranded oligonucleotide comprising;

a first nucleic acid molecule having a first nucleotide sequence that is antisense to the target gene and having 2′

-O-methyl RNA arms and an unmodified DNA gap, and

a second nucleic acid molecule having a nucleotide sequence that hybridizes to the first nucleotide sequence,

wherein the first and second molecules are each 25 nucleosides in length and are 100% complementary to each other,

wherein the double-stranded oligonucleotide has no overhanging single-stranded sequence at either end of the molecule,

to thereby decrease expression of the target gene in the cell.

2. A method of decreasing expression of a target gene in a cell, comprising:

contacting the cell with a composition comprising a double-stranded oligonucleotide in the presence of a transfection reagent wherein the transfection reagent is a cationic lipid, the double-stranded oligonucleotide comprising;

a first nucleic acid molecule having a first nucleotide sequence that is antisense to the target gene and having 2′

-O-methyl RNA arms and an unmodified DNA gap, and

a second nucleic acid molecule having a nucleotide sequence that hybridizes to the first nucleotide sequence,

wherein the first and second molecules are each 25 nucleosides in length and are 100% complementary to each other,

wherein the double-stranded oligonucleotide has no overhanging single-stranded sequence at either end of the molecule,

to thereby decrease expression of the target gene in the cell.

3. A method of decreasing expression of a target gene in a cell, comprising:

contacting the cell with a composition comprising a double-stranded oligonucleotide in the presence of a transfection reagent wherein the transfection reagent is a cationic lipid, the double-stranded oligonucleotide comprising;

a first nucleic acid molecule having a first nucleotide sequence that is antisense to the target gene and having 2′

-O-methyl RNA arms and an unmodified DNA gap, and

a second nucleic acid molecule having a nucleotide sequence that hybridizes to the first nucleotide sequence,

wherein the first and second molecules are each 25 nucleosides in length to thereby decrease expression of the target gene in the cell.