Immunostimulatory nucleic acid molecules

• US 20050277609A1
• Filed: 01/07/2005
• Published: 12/15/2005
• Est. Priority Date: 07/15/1994
• Status: Active Grant

• Current Assignee: Coley Pharmaceutical Group, Inc. (Pfizer Inc.), United States Department Of Health And Human Services, University of Iowa Research Foundation
• Original Assignee: Coley Pharmaceutical Group, Inc. (Pfizer Inc.), The United States of America As Represented By The Secretary of Agriculture, University of Iowa Research Foundation
• Inventors: Krieg, Arthur M., Klinman, Dennis, Steinberg, Alfred D., Kline, Joel
• Granted Patent: US 8,158,592 B2
• Time in Patent Office: Days
• US Class Current: 514/44.A
• 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. Methods for treating atopic diseases, including atopic dermatitis, are disclosed.

Claims

1-18. -18. (canceled)

19. An immunostimulatory oligonucleotide of at least 8 nucleotides in length comprising a sequence selected from the group consisting of AACGTC and GGCGTT.

39. A method of modulating an immune response in a subject comprising administering the immunostimulatory oligonucleotide of claim 19 to a subject in an amount sufficient to modulate the immune response.

51. A method according to claim 39, wherein the subject is suffering from a disorder selected from the group consisting of cancer, allergic disease, asthma and an infectious disease.

52. A method according to claim 51, wherein the infectious disease is caused by a virus selected from the group consisting of hepatitis B virus, papillomavirus and human immunodeficiency virus.

23. An immunostimulatory composition comprising an immunostimulatory nucleic acid and an antigen, wherein the immunostimulatory nucleic acid comprises 5′

-cytosine, guanine-3′

, and wherein the immunostimulatory nucleic acid and the antigen and are proximately associated at a distance effective to enhance an immune response.

24. The immunostimulatory composition of claim 23, wherein the immunostimulatory nucleic acid comprises a palindromic region, and wherein the palindromic region comprises the sequence 5′

-cytosine, guanine-3′

.

25. The immunostimulatory composition of claim 24, wherein the immunostimulatory nucleic acid comprises 5′

-purine, purine, cytosine, guanine, pyrimidine, pyrimidine-3′

.

26. The immunostimulatory composition of claim 25, wherein the immunostimulatory nucleic acid comprises a sequence selected from the group consisting of AACGTT, AGCGTT, GACGTT, GGCGTT, AACGTC, and AACGCT.

27. The immunostimulatory composition of claim 25, wherein the immunostimulatory nucleic acid is selected from the group consisting of AACGTC and GGCGTT.

28. The immunostimulatory composition of claim 23, wherein the immunostimulatory nucleic acid and antigen are proximately associated by encapsulation.

29. The immunostimulatory composition of claim 28, wherein the encapsulation is within liposomes.

30. The immunostimulatory composition of claim 23, wherein the immunostimulatory nucleic acid and antigen are proximately associated by linkage to a delivery molecule.

31. The immunostimulatory composition of claim 23, wherein the immunostimulatory nucleic acid and antigen are proximately associated such that the immunostimulatory nucleic acid and the antigen are co-delivered to an immune target.

32. The immunostimulatory composition of claim 31, wherein the immune target is an antigen presenting cell.

33. The immunostimulatory composition of claim 32, wherein the antigen presenting cell is a dendritic cell.

34. The immunostimulatory composition of claim 32, wherein the antigen presenting cell is a macrophage cell.

35. The immunostimulatory composition of claim 32, wherein the antigen presenting cell is a lymphocyte.

36. The immunostimulatory composition of claim 23, further comprising an adjuvant.

37. The immunostimulatory composition of claim 35, wherein the immunostimulatory nucleic acid and antigen are proximately associated by encapsulation.

38. The immunostimulatory composition of claim 35, wherein the immunostimulatory nucleic acid and antigen are proximately associated by linkage to a delivery molecule.

45. A method of modulating an immune response in a subject comprising the administration of an immunostimulatory composition according to claim 23 in an amount sufficient to modulate the immune response.

46. The method of claim 45, wherein the modulating of an immune response comprises induction of a Th1 response.

47. A method of modulating an immune response in a subject comprising the administration of an immunostimulatory composition according to claim 27 in an amount sufficient to modulate the immune response.

48. The method of claim 47, wherein the modulating of an immune response comprises induction of a Th1 response.

49. A method of modulating an immune response in a subject comprising the administration of an immunostimulatory composition according to claim 36 in an amount sufficient to modulate the immune response.

50. The method of claim 49, wherein the modulating of an immune response comprises induction of a Th1 response.

60. A method of preventing an infectious disease in a subject comprising administration of an immunostimulatory composition according to claim 23.

61. A method of preventing an infectious disease in a subject comprising administration of an immunostimulatory composition according to claim 27.

62. A method to screen for human immunostimulatory activity of oligonucleotides comprising the steps of:

(a) providing macrophage cells and an oligonucleotide to be tested;

(b) incubating the cells and oligonucleotide of step a) for an appropriate length of time;

(c) determining the relative amount of Th1-biased cytokines in the cell culture supernatant.

63. A method to screen for human immunostimulatory activity of oligonucleotides according to claim 62, wherein at least one of the Th1-biased cytokines determined is interferon-gamma.

64. A method to screen for human immunostimulatory activity of oligonucleotides according to claim 62, wherein at least one of the Th1-biased cytokines determined is interleukin-12.

65. An immunostimulatory composition comprising (a) an immunostimulatory nucleic acid;

(b) an antigen; and

(c) an adjuvant other than alum, wherein the immunostimulatory nucleic acid comprises 5′

-cytosine, guanine-3′

.

66. The immunostimulatory composition of claim 65, wherein the immunostimulatory nucleic acid comprises a palindromic region, and wherein the palindromic region comprises the sequence 5′

-cytosine, guanine-3′

.

67. A method of modulating an immune response in a subject, comprising administering the composition of claim 65 in an amount sufficient to modulate the immune response.

68. An immunostimulatory oligonucleotide of at least 8 nucleotides in length represented by the formula:

69. The immunostimulatory oligonucleotide of claim 68, wherein the oligonucleotide includes more than one CpG motif.

70. An immunostimulatory oligonucleotide of at least 8 nucleotides in length represented by the formula:

71. The immunostimulatory oligonucleotide of claim 70, wherein X₁X₂ is selected from the group consisting of GpT, GpG, GpA and ApA and X₃X₄ is selected from the group consisting of TpT, CpT and GpT.

72. The immunostimulatory oligonucleotide of claim 70, wherein X₁X₂ is GpG, GpA or ApA and X₃X₄ is TpT or CpT.

73. The immunostimulatory oligonucleotide of claim 70, wherein the oligonucleotide includes more than one CpG motif.