Immunostimulatory nucleic acid molecules

• US 6,207,646 B1
• Filed: 10/30/1996
• Issued: 03/27/2001
• Est. Priority Date: 07/15/1994
• Status: Expired due to Term

• Current Assignee: United States As Represented By The Secretary Department of Health and Human Services, University of Iowa Research Foundation
• Original Assignee: Coley Pharmaceutical Group, Inc. (Pfizer Inc.), United States Department Of Health And Human Services, University of Iowa Research Foundation
• Inventors: Kline, Joel, Krieg, Arthur M., Klinman, Dennis, Steinberg, Alfred D.
• Primary Examiner(s): Martinell, James
• Application Number:
  US08/738,652
• Time in Patent Office: 1,609 Days
• Field of Search: 514/44, 536/23.1, 435/320.1
• US Class Current: 514/44.00R
• CPC Class Codes: A61K 2039/55561 CpG containing adjuvants; Oligonucleotide containing adjuvants
• CPC Class Codes: A61K 31/00 Medicinal preparations containing organic active ingredients
• CPC Class Codes: A61K 31/4706 4-Aminoquinolines; 8-Aminoquinolines, e.g. chloroquine, primaquine
• CPC Class Codes: A61K 31/7048 having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin , digitoxin or digoxin
• CPC Class Codes: A61K 31/711 Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
• CPC Class Codes: A61K 31/7125 Nucleic acids or oligonucleotides having modified internucleoside linkage, i.e. other than 3'-5' phosphodiesters
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/02 Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
• 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 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 17/06 Antipsoriatics
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 31/10 Antimycotics
• 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 37/02 : 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: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• 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 2310/315 : Phosphorothioates
• CPC Class Codes: C12Q 1/68 : involving nucleic acids
• 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

Nucleic acids containing unmethylated CpG dinucleotides and therapeutic utilities based on their ability to stimulate an immune response and to redirect a Th2 response to a Th1 response in a subject are disclosed.

Claims

1. A method for ameliorating an immune system deficiency in a subject, comprising the steps of:

a) contacting lymphocytes obtained from the subject with an antigen and an immunostimulatory nucleic acid, comprising;

2. A method of claim 1, wherein the immune system deficiency is selected from the group consisting of:

the presence of a tumor, cancer or infectious agent in the subject.

3. A method for desensitizing a subject against the occurrence of an allergic reaction in response to contact with a particular allergen, comprising administering to the subject an effective amount of an immunostimulatory nucleic acid, comprising:

4. A method of vaccination in a subject, comprising administering to the subject a vaccine antigen or an antigen encoded in a DNA vaccine and immunostimulatory nucleic acid, comprising:

5. A method for treating leukemia in a subject, comprising administering to the subject an immunostimulatory nucleic acid, comprising:

6. A composition comprising:

a plasmid including an immunostimulatory nucleic acid sequence, comprising;

7. The composition of claim 6,

wherein X₁X₂ are nucleotides selected from the group consisting of:

GpT, GpG, GpA and ApA; and

X₃X₄ are nucleotides selected from the group consisting of;

TpT, CpT, GpT, and TpG.

8. The composition of claim 6, further comprising a B-cell targeting molecule.

9. The composition of claim 8, wherein the targeting molecule is selected from the group consisting of a sterol, a lipid and a B-cell specific binding agent.

10. The composition of claim 6, wherein the antigen is encoded in a DNA vaccine.

11. The composition of claim 6, wherein the antigen is selected from the group consisting of proteins, polysaccharides, polysaccharide conjugates, glycolipids, viruses, bacteria, fungi, parasites, and allergens.

12. The composition of claim 11, wherein the antigen is an allergen.

13. The composition of claim 11, wherein the antigen is derived from an infectious organism selected from the group consisting of infectious bacteria, infectious virus, and infectious fungi.

14. A composition comprising:

an isolated immunostimulatory nucleic acid of 8 to 100 nucleotides in length, comprising;

15. The composition of claim 14, wherein the isolated immunostimulatory nucleic acid includes a phosphate backbone modification which is a phosphorothioate or phosphorodithioate modification.

16. The composition of claim 14, wherein the antigen is encoded in a DNA vaccine.

17. The composition of claim 14, wherein the antigen is selected from the group consisting of proteins, polysaccharides, polysaccharide conjugates, glycolipids, viruses, bacteria, fungi, parasites, and allergens.

26. A method of inducing an antigen-specific immune response in a subject comprising:

administering a vaccine to a subject, wherein the vaccine includes an antigen in combination with an immunostimulatory nucleic acid of claims 6, 14, 18 or 22 in an amount effective to induce an immune response.

27. The method of claim 26, wherein the antigen is selected from the group consisting of proteins, polysaccharides or polysaccharide conjugates, glycolipids, viruses, bacteria, fungi, parasites, and allergens.

28. The method of claim 27, wherein leukocytes of the subject are isolated and contacted with the antigen and immunostimulatory nucleic acid to produce activated leukocytes and wherein the activated leukocytes are readministered to the subject.

29. The method of claim 26, wherein the vaccine is administered ex vivo.

18. A composition comprising:

an immunostimulatory nucleic acid comprising;

19. The composition of claim 18, wherein the synthetic immunostimulatory nucleic acid includes a phosphate backbone modification which is a phosphorothioate or phosphorodithioate modification.

20. The composition of claim 18, wherein the antigen is encoded in a DNA vaccine.

21. The composition of claim 18, wherein the antigen is selected from the group consisting of proteins, polysaccharides, polysaccharide conjugates, glycolipids, viruses, bacteria, fungi, parasites, and allergens.

30. The composition of claim 18, wherein the immunostimulatory nucleic acid molecule is 8 to 100 nucleotides in length.

22. A composition comprising:

an immunostimulatory nucleic acid of 8 to 40 nucleotides in length, comprising;

23. The composition of claim 22, wherein the immunostimulatory nucleic acid includes a phosphate backbone modification which is a phosphorothioate or phosphorodithioate modification.

24. The composition of claim 22, wherein the antigen is encoded in a DNA vaccine.

25. The composition of claim 22, wherein the antigen is selected from the group consisting of proteins, polysaccharides, polysaccharide conjugates, glycolipids, viruses, bacteria, fungi, parasites, and allergens.

31. A composition comprising:

an immunostimulatory nucleic acid, comprising;

32. The composition of claim 31, wherein the immunostimulatory nucleic acid molecule is 8 to 100 nucleotides in length.

33. The composition of claim 31, wherein X₁X₂ are dinucleotides selected from the group consisting of:

GpT, GpG, GpA ApA, ApT, ApG, CpT, CpA, CpG, TpA, TpT, and TpG.

34. The composition of claim 31, wherein X₃X₄ are dinucleotides selected from the group consisting of:

TpT, CpT, ApT, TpG, ApG, CpG, TpC, ApC, CpC, TpA, ApA, and CpA.

35. The composition of claim 31, wherein the immunostimulatory nucleic acid is associated with a cationinc lipid.

36. The composition of claim 31, wherein X₁X₂ are dinucleotides selected from the group consisting of GpT, GpG, GpA, and ApA and wherein X₃X₄ are dinucleotides selected from the group consisting of TpT, CpT and ApT.

37. The composition of claim 31, wherein the antigen is selected from the group consisting of proteins, polysaccharides, polysaccharide conjugates, glycolipids, viruses, bacteria, fungi, parasites, and allergens.

38. The composition of claim 37, wherein the antigen is an allergen.

39. The composition of claim 37, wherein the antigen is derived from an infectious organism selected from the group consisting of infectious bacteria, infectious virus, and infectious fungi.