Oligonucleotide compositions with enhanced efficiency

• US 10,626,398 B2
• Filed: 01/21/2019
• Issued: 04/21/2020
• 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, Taylor, Margaret
• Primary Examiner(s): McGarry, Sean
• Application Number:
  US16/253,180
• Publication Number:
  US 20190316138A1
• Time in Patent Office: 456 Days
• Field of Search: None
• 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 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 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/315 Phosphorothioates
• CPC Class Codes: C12N 2320/11 : for the determination of target sites, i.e. of active nucleic acids
• CPC Class Codes: C12N 2320/30 : Special therapeutic applications
• CPC Class Codes: C12N 2320/31 : Combination therapy
• CPC Class Codes: C12N 2320/50 : Methods for regulating/modulating their activity
• CPC Class Codes: C12N 2330/30 : chemically synthesised
• 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

The oligonucleotide compositions of the present invention make use of combinations of oligonucleotides. In one aspect, the invention features an oligonucleotide composition including at least 2 different oligonucleotides targeted to a target gene. This invention also provides methods of inhibiting protein synthesis in a cell and methods of identifying oligonucleotide compositions that inhibit synthesis of a protein in a cell.

Claims

1. A composition comprising a combination of double-stranded oligonucleotides, the combination consisting of three or more different double-stranded oligonucleotides in equimolar concentrations,

wherein each double-stranded oligonucleotide is targeted to a different sequence within a single target gene,

each double-stranded oligonucleotide consists of two separate strands,

each strand is between 15 and 40 nucleomonomers in length;

wherein the combination is capable of RNA interference (RNAi).

2. The composition of claim 1 wherein each strand is between 20 and 30 nucleomonomers in length.

3. The composition of claim 1 wherein at least one of the oligonucleotides comprises at least one modified sugar moiety or at least one modified internucleoside linkage.

4. The composition of claim 1 wherein each double-stranded oligonucleotide comprises a nucleotide sequence that is at least 90% identical to a portion of the target gene.

5. The composition of claim 1 wherein at least one of the double-stranded oligonucleotides is covalently linked to an agent to facilitate cellular uptake.

6. The composition of claim 1, wherein the target gene is a mammalian gene.

20. A method of inhibiting protein synthesis in a cell comprising

contacting the cell with the composition of claim 1 thereby inhibiting the synthesis of a target protein.

7. A composition comprising a combination of double-stranded oligonucleotides, the combination consisting of two different double-stranded oligonucleotides in equimolar concentrations,

wherein each double-stranded oligonucleotide is targeted to a different sequence within a single target gene,

each double-stranded oligonucleotide consists of two separate strands,

each strand is between 15 and 40 nucleomonomers in length,

wherein the combination is capable of RNA interference (RNAi).

8. The composition of claim 7 wherein each strand is between 20 and 30 nucleomonomers in length.

9. The composition of claim 7 wherein at least one of the oligonucleotides comprises at least one modified sugar moiety or at least one modified internucleoside linkage.

10. The composition of claim 7 wherein each double-stranded oligonucleotide comprises a nucleotide sequence that is at least 90% identical to a portion of the target gene.

11. The composition of claim 7 wherein at least one of the double-stranded oligonucleotides is covalently linked to an agent to facilitate cellular uptake.

12. The composition of claim 7, wherein the target gene is a mammalian gene.

21. A method of inhibiting protein synthesis in a cell comprising

contacting the cell with the composition of claim 7 thereby inhibiting the synthesis of a target protein.

13. A composition comprising a combination of double-stranded oligonucleotides, the combination consisting of three, four, five, six, seven or eight different double-stranded oligonucleotides in equimolar concentration,

wherein each double-stranded oligonucleotide is targeted to a different sequence within a single target gene,

each double-stranded oligonucleotide consists of two separate strands,

each strand is between 15 and 40 nucleomonomers in length,

wherein the combination is capable of RNA interference (RNAi).

14. The composition of claim 13 wherein each strand is between 20 and 30 nucleomonomers in length.

15. The composition of claim 13 wherein at least one of the oligonucleotides comprises at least one modified sugar moiety or at least one modified internucleoside linkage.

16. The composition of claim 13 wherein each double-stranded oligonucleotide comprises a nucleotide sequence that is at least 90% identical to a portion of the target gene.

17. The composition of claim 13 wherein at least one of the double-stranded oligonucleotides is covalently linked to an agent to facilitate cellular uptake.

18. The composition of claim 13 wherein the combination of double-stranded oligonucleotides consists of three or four different double-stranded oligonucleotides in equimolar concentration.

19. The composition of claim 18 wherein the combination of double-stranded oligonucleotides consists of four different double-stranded oligonucleotides in equimolar concentration.

22. A method of inhibiting protein synthesis in a cell comprising

contacting the cell with the composition of claim 13 thereby inhibiting the synthesis of a target protein.