Double-stranded oligonucleotides

• US 10,106,793 B2
• Filed: 01/27/2017
• Issued: 10/23/2018
• 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, Wiederholt, Kristin
• Primary Examiner(s): Whiteman, Brian A
• Application Number:
  US15/418,653
• Publication Number:
  US 20170137816A1
• Time in Patent Office: 634 Days
• 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 composition comprising a double-stranded oligonucleotide molecule, the molecule comprising:

a first strand and a second strand each having a length of less than 25 nucleotides and having a duplex length of at least 10 to at least 24 nucleotides;

wherein at least one but not all of the nucleotides of the first strand comprises a 2′

-O-methyl modification and at least one but not all of the nucleotides of the second strand comprises a 2′

-O-methyl modification, and

wherein said double-stranded oligonucleotide molecule is chemically synthesized.

2. The composition of claim 1, wherein a strand of said double-stranded oligonucleotide molecule is complementary to a sequence of a target nucleic acid.

3. The composition of claim 2 wherein the target nucleic acid is a mRNA or a gene.

4. The composition of claim 2 wherein the target nucleic acid is present within a cell.

5. The composition of claim 4 wherein the cell is a eukaryotic cell.

6. The composition of claim 1 wherein the double-stranded oligonucleotide molecule is a double-stranded ribonucleic acid molecule.

7. The composition of claim 1 wherein the duplex length is at least 15 nucleotides.

8. A method for introducing a double-stranded ribonucleic acid molecule comprising a first strand and a second strand into a eukaryotic cell in vitro, the method comprising contacting the eukaryotic cell with the double-stranded ribonucleic acid molecule,

wherein the first strand and the second strand each have a length of less than 25 nucleotides and have a duplex length of at least 10 to at least 24 nucleotides,

wherein at least one but not all of the nucleotides of the first strand of the double-stranded ribonucleic acid molecule comprises a 2′

-O-methyl modification;

wherein at least one but not all of the nucleotides of the second strand of the double-stranded ribonucleic acid molecule comprises a 2′

-O-methyl modification; and

wherein said double-stranded ribonucleic acid molecule is chemically synthesized;

to result in the double-stranded nucleic acid molecule being introduced into the eukaryotic cell.

9. The method of claim 8, wherein the eukaryotic cell is contacted with the double-stranded ribonucleic acid molecule in the presence of a transfection agent.

10. The method of claim 9, wherein the transfection reagent is a cationic lipid.

11. The method of claim 8, wherein the double-stranded ribonucleic acid molecule is introduced into the cell by electroporation.

12. The method of claim 8, wherein a strand of said double-stranded ribonucleic acid molecule is complementary to a sequence of an mRNA expressed in said eukaryotic cell.

13. The method of claim 8, wherein a strand of the double-stranded ribonucleic acid molecule has at least 80% sequence complementarity with an mRNA expressed in the eukaryotic cell.

14. The method of claim 8, wherein a strand of the double-stranded ribonucleic acid molecule has at least 90% sequence complementarity with an mRNA expressed in the eukaryotic cell.

15. The method of claim 8, wherein a strand of the double-stranded ribonucleic acid molecule has at least 95% sequence complementarity with an mRNA expressed in the eukaryotic cell.

16. The method of claim 12 wherein introducing the double-stranded ribonucleic acid molecule into the eukaryotic cell inhibits gene expression in the cell.

17. The method of claim 8 wherein the duplex length is at least 15 nucleotides.