STRUCTURE AND USE OF 5' PHOSPHATE OLIGONUCLEOTIDES

• US 20120288476A1
• Filed: 05/08/2012
• Published: 11/15/2012
• Est. Priority Date: 08/08/2006
• Status: Active Grant

• Current Assignee: Rheinische Friedrich-Wilhelms Universitã¤T Bonn
• Original Assignee: Rheinische Friedrich-Wilhelms Universitã¤T Bonn
• Inventors: Hartmann, Gunther, Hornung, Veit
• Granted Patent: US 9,381,208 B2
• Time in Patent Office: Days
• US Class Current: 424/85.4
• CPC Class Codes: A61K 31/7105 Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 31/7115 Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 33/00 Antiparasitic agents
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 37/08 Antiallergic agents antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12N 15/117 Nucleic acids having immunomodulatory properties, e.g. containing CpG-motifs
• CPC Class Codes: C12N 2310/17 Immunomodulatory nucleic acids
• CPC Class Codes: C12N 2310/31 of the backbone
• CPC Class Codes: C12N 2310/32 of the sugar
• CPC Class Codes: C12N 2310/335 : Modified T or U
• CPC Class Codes: C12N 2310/346 : having a combination of backbone and sugar modifications
• CPC Class Codes: C12N 2310/351 : Conjugate
• CPC Class Codes: C12N 2320/30 : Special therapeutic applications
• 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

Oligonucleotides bearing free, uncapped 5′ phosphate group(s) are recognized by RIG-I, leading to the induction of type I IFN, IL-18 and IL-1β production. Bacterial RNA also induces type I IFN production. 5′ phosphate oligonucleotides and bacterial RNA can be used for inducing an anti-viral response or an anti-bacterial response, in particular, type I IFN and/or IL-18 and/or IL-1β production, in vitro and in vivo and for treating various disorders and diseases such as viral infections, bacterial infections, parasitic infections, tumors, allergies, autoimmune diseases, immunodeficiencies and immunosuppression. Single-stranded 5′ triphosphate RNA can be used for inducing an anti-viral response, an anti-bacterial response, or an anti-tumor response, in particular, type I IFN and/or IL-18 and/or IL-1β production, in a target cell-specific manner.

Claims

1. An oligonucleotide wherein the oligonucleotide

(a) is at least 12 nucleotides in length;

(b) comprises a 5′

triphosphate; and

(c) comprises at least 1 ribonucleotide at the 5′

end, wherein the first ribonucleotide at the 5′

end comprises a G, U or C.

2. The oligonucleotide of claim 1, wherein the oligonucleotide contains a 2′

-methyl-dNTP and/or 2′

-fluorine-dNTP modification.

3. The oligonucleotide of claim 1, wherein the oligonucleotide is covalently linked to at least one antigen.

4. A pharmaceutical composition comprising the oligonucleotide of claim 1 or a viral vector encoding the oligonucleotide of claim 1.

5. The pharmaceutical composition of claim 4, further comprising one or more of a complexation agent, an antiviral agent, an anti-bacterial agent, an anti-tumor agent, an immunostimulatory agent, retinoic acid, type I IFN, and an antigen.

6. A method of preventing or treating a disease or disorder in a vertebrate animal comprising administering an effective amount of the pharmaceutical composition of claim 4 to the vertebrate animal in need thereof, thereby preventing or treating a disease or disorder in the vertebrate animal.

7. The method of claim 6, wherein the disease or disorder is a viral infection, a bacterial infection, an immunosuppression, an immunodeficiency, a tumor, or an autoimmune disease.

8. The method of claim 6, wherein the pharmaceutical composition stimulates an immune response in the vertebrate animal.

9. The method of claim 8, wherein the immune response is an anti-viral or anti-bacterial response.

10. The method of claim 9, wherein the anti-viral or anti-bacterial response comprises type I IFN production, IL-18 production, and/or IL-1β

production.

11. The method of claim 6, further comprising administering one or more of a complexation agent, an antiviral agent, an anti-bacterial agent, an anti-tumor agent, anti-tumor therapy, an immunostimulatory agent, retinoic acid, type I IFN, and an antigen to the vertebrate animal.

12. A method for inducing apoptosis of a tumor cell, comprising contacting a tumor cell with the pharmaceutical composition of claim 4 to thereby induce apoptosis of the tumor cell.

13. An in vitro method for stimulating an anti-viral or an anti-bacterial response in a cell, comprising the steps of:

(a) mixing the oligonucleotide of claim 1 with a complexation agent; and

(b) contacting a cell with the mixture of (a), wherein the cell expresses RIG-I and/or NALP-3.

14. The in vitro method of claim 13, wherein the anti-viral or anti-bacterial response comprises type I IFN production, IL-1β

production, and/or IL-1 production.