Methods related to immunostimulatory nucleic acid-induced interferon

• US 6,949,520 B1
• Filed: 09/27/2000
• Issued: 09/27/2005
• Est. Priority Date: 09/27/1999
• Status: Expired due to Fees

• Current Assignee: Coley Pharmaceutical GmbH (Pfizer Inc.), University of Iowa Research Foundation
• Original Assignee: Coley Pharmaceutical GmbH (Pfizer Inc.), Coley Pharmaceutical Group, Inc. (Pfizer Inc.), University of Iowa Research Foundation
• Inventors: Krieg, Arthur M., Hartmann, Gunther, Bratzler, Robert L.
• Primary Examiner(s): Guzo, David
• Assistant Examiner(s): Nguyen, Quang
• Application Number:
  US09/672,126
• Time in Patent Office: 1,826 Days
• Field of Search: 514/44, 435/455, 435/375, 424/85.7
• US Class Current: 514/44.00R
• CPC Class Codes: A61K 2039/5154 Antigen presenting cells [APCs], e.g. dendritic cells or macrophages
• CPC Class Codes: A61K 2039/5158 Antigen-pulsed cells, e.g. T-cells
• CPC Class Codes: A61K 2039/55561 CpG containing adjuvants; Oligonucleotide containing adjuvants
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 38/21 Interferons [IFN]
• CPC Class Codes: A61K 39/461 characterised by the cell type used
• CPC Class Codes: A61K 39/464411 Immunoglobulin superfamily
• CPC Class Codes: A61K 39/464419 Receptors for interleukins [IL]
• CPC Class Codes: A61K 39/464463 Phosphatases
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 31/20 for DNA viruses
• CPC Class Codes: A61P 31/22 for herpes viruses
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12N 15/117 Nucleic acids having immunomodulatory properties, e.g. containing CpG-motifs
• CPC Class Codes: C12N 2501/056 Immunostimulating oligonucleotides, e.g. CpG
• CPC Class Codes: C12N 2501/22 Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2501/23 Interleukins [IL]
• CPC Class Codes: C12N 5/0639 Dendritic cells, e.g. Langherhans cells in the epidermis

Abstract

Methods and compositions are provided for extending the clinical utility of IFN-α in the treatment of a variety of viral and proliferative disorders. Among other aspects, the invention provides methods which increase the efficacy of IFN-α treatment and reduce IFN-α treatment-related side effects. In addition, methods are provided for supporting the survival and for activating natural interferon producing cells (IPCs) in vitro without exogenous IL-3 or GM-CSF. The invention is based on the discovery that certain CpG and non-CpG ISNAs promote survival and stimulation of IPCs.

Claims

1. In a method which calls for administration of interferon alpha (IFN-α

) to a mammalian subject, the improvement comprising co-administering to the mammalian subject an effective amount of an isolated immunostimulatory nucleic acid, wherein said isolated immunostimulatory nucleic acid is 10 to 100 nucleotides long and comprises a poly-G sequence at each end and a central palindromic sequence comprising an unmethylated CpG dinucleotide.

2. The improvement of claim 1, wherein the IFN-α

is administered at a dose below the clinically established effective dose for IFN-α

alone.

3. The improvement of claim 1, wherein the IFN-α

is administered at the maximum tolerated dose for IFN-α

in the absence of the nucleic acid.

4. The improvement of claim 1, wherein the IFN-α

is administered at least 20 percent below the maximum tolerated dose of IFN-α

in the subject.

5. The improvement of claim 1, wherein the IFN-α

is administered at least 30 percent below the maximum tolerated dose of IFN-α

in the subject.

6. The improvement of claim 1, wherein the IFN-α

is administered at least 40 percent below the maximum tolerated dose of IFN-α

in the subject.

7. The improvement of claim 1, wherein the IFN-α

is administered at least 50 percent below the maximum tolerated dose of IFN-α

in the subject.

8. The improvement of claim 1, wherein the immunostimulatory nucleic acid is stabilized.

9. The improvement of claim 1, wherein the immunostimulatory nucleic acid comprises a backbone with at least one nuclease-resistant internucleotide linkage selected from the group consisting of:

phosphorothioate, phosphorodithioate, methylphosphonate, and peptide.

10. The improvement of claim 1, wherein the immunostimulatory nucleic acid comprises at least one nucleotide analog or derivative.

11. The improvement of claim 1, wherein the immunostimulatory nucleic acid is between 12 and 40 nucleotides in length.

12. The improvement of claim 1, wherein the immunostimulatory nucleic acid has a sequence selected from the group consisting of

ggGGGACGATCGTCgggggGODN 2216SEQ ID NO:

13. The improvement of claim 1, further comprising co-administering GM-CSF to the subject.

14. The improvement of claim 1, wherein the subject has a condition selected from the group consisting of a proliferative disorder and a viral infection.

15. The improvement of claim 1, wherein the subject has a proliferative disorder selected from the group consisting of:

hairy cell leukemia, chronic myelogenous leukemia, cutaneous T-cell leukemia, multiple myeloma, follicular lymphoma, malignant melanoma, squamous cell carcinoma, AIDS-related Kaposi'"'"'s sarcoma, renal cell carcinoma, prostate carcinoma, bladder cell carcinoma, cervical dysplasia, and colon carcinoma.

16. The improvement of claim 1, wherein the subject has a viral infection selected from the group consisting of:

hepatitis B, hepatitis C, condyloma acuminatum, human immunodeficiency virus, herpes, cytomegalovirus, Epstein-Barr virus, and papillomavirus.

24. The method of claim 1, wherein the co-administering comprises administering the IFN-α

and the isolated immunostimulatory nucleic acid together.

25. The method of claim 1, wherein the co-administering comprises administering the IFN-α

and the isolated immunostimulatory nucleic acid sequentially.

17. A method of supplementing interferon alpha (IFN-α

) treatment of a subject, comprising

administering to a mammalian subject in need of IFN-α

treatment an effective amount of IFN-α and

an isolated immunostimulatory nucleic acid, wherein said isolated immunostimulatory nucleic acid is 10 to 100 nucleotides long and comprises a poly-G sequence at each end and a central palindromic sequence comprising an unmethylated CpG dinucleotide.

26. The method of claim 17, wherein the IFN-α

is administered at a dose below a clinically established effective dose for IFN-α

alone.

27. The method of claim 17, wherein the IFN-α

is administered at a maximum tolerated dose for IFN-α

in absence of the immunostimulatory nucleic acid.

28. The method of claim 17, wherein the IFN-α

is administered at least 20 percent below a maximum tolerated dose of IFN-α

in the subject.

29. The method of claim 17, wherein the IFN-α

is administered at least 30 percent below a maximum tolerated dose of IFN-α

in the subject.

30. The method of claim 17, wherein the IFN-α

is administered at least 40 percent below a maximum tolerated dose of IFN-α

in the subject.

31. The method of claim 17, wherein the IFN-α

is administered at least 50 percent below a maximum tolerated dose of IFN-α

in the subject.

32. The method of claim 17, wherein the immunostimulatory nucleic acid is stabilized.

33. The method of claim 17, wherein the immunostimulatory nucleic acid comprises a backbone with at least one nuclease-resistant internucleotide linkage selected from the group consisting of:

phosphorothioate, phosphorodithioate, methylphosphonate, and peptide.

34. The method of claim 17, wherein the immunostimulatory nucleic acid comprises at least one nucleotide analog or derivative.

35. The method of claim 17, herein the immunostimulatory nucleic acid is between 12 and 40 nucleotides in length.

36. The method of claim 17, wherein the immunostimulatory nucleic acid has a sequence selected from the group consisting of

ggGGGACGATCGTCgggggGODN 2216SEQ ID NO:

37. The method of claim 17, further comprising co-administering GM-CSF to the subject.

38. The method of claim 17, wherein the subject has a condition selected from the group consisting of a proliferative disorder and a viral infection.

39. The method of claim 17, wherein the subject has a proliferative disorder selected from the group consisting of:

hairy cell leukemia, chronic myelogenous leukemia, cutaneous T-cell leukemia, multiple myeloma, follicular lymphoma, malignant melanoma, squamous cell carcinoma, AIDS-related Kaposi'"'"'s sarcoma, renal cell carcinoma, prostate carcinoma, bladder cell carcinoma, cervical dysplasia, and colon carcinoma.

40. The method of claim 17, wherein the subject has a viral infection selected from the group consisting of:

hepatitis B, hepatitis C, condyloma acuminatum, human immunodeficiency virus, herpes, cytomegalovirus, Epstein-Barr virus, and papillomavirus.

18. A method of increasing efficacy of interferon alpha (IFN-α

) treatment of a subject, comprising;

administering to a mammalian subject in need of treatment with IFN-α

a pharmaceutical composition comprising IFN-α

, and

coadministering to the subject in need of such treatment a pharmaceutical composition comprising an immunostimulatory nucleic acid in an amount which, together with the administered IFN-α

, is an effective IFN-α

treatment, wherein the efficacy of the IFN-α

treatment is greater than the efficacy of administering the same amount of IFN-α

in the absence of coadministering the immunostimulatory nucleic acid, and wherein said immunostimulatory nucleic acid is 10 to 100 nucleotides long and comprises a poly-G sequence at each end and a central palindromic sequence comprising an unmethylated CpG dinucleotide.

41. The method of claim 18, wherein the coadministering comprises administering the IFN-α

and the immunostimulatory nucleic acid together.

42. The method of claim 18, wherein the coadministering comprises administering the IFN-α

and the immunostimulatory nucleic acid sequentially.

43. The method of claim 18, wherein the pharmaceutical composition comprising an immunostimulatory nucleic acid is administered locally.

44. The method of claim 18, wherein the immunostimulatory nucleic acid is stabilized.

45. The method of claim 18, wherein the immunostimulatory nucleic acid comprises a backbone with at least one nuclease-resistant internucleotide linkage selected from the group consisting of:

phosphorothioate, phosphorodithioate, methylphosphonate, and peptide.

46. The method of claim 18, wherein the immunostimulatory nucleic acid comprises at least one nucleotide analog or derivative.

47. The method of claim 18, wherein the immunostimulatory nucleic acid is between 12 and 40 nucleotides in length.

48. The method of claim 18, wherein the immunostimulatory nucleic acid has a sequence selected from the group consisting of

ggGGGACGATCGTCgggggGODN 2216SEQ ID NO:

49. The method of claim 18, wherein the subject has a condition selected from the group consisting of a proliferative disorder and a viral infection.

50. The method of claim 18, wherein the subject has a proliferative disorder selected from the group consisting of:

hairy cell leukemia, chronic myelogenous leukemia, cutaneous T-cell leukemia, multiple myeloma, follicular lymphoma, malignant melanoma, squamous cell carcinoma, AIDS-related Kaposi'"'"'s sarcoma, renal cell carcinoma, prostate carcinoma, bladder cell carcinoma, cervical dysplasia, and colon carcinoma.

51. The method of claim 18, wherein the subject has a viral infection selected from the group consisting of:

hepatitis B, hepatitis C, condyloma acuminatum, human immunodeficiency virus, herpes, cytomegalovirus, Epstein-Barr virus, and papillomavirus.

19. A method of decreasing a dose of interferon alpha (IFN-α

) effective for treating a subject, comprising;

administering to a mammalian subject in need of treatment with IFN-α

a pharmaceutical composition comprising IFN-α

, and

coadministering to the subject in need of such treatment a pharmaceutical composition comprising an immunostimulatory nucleic acid in an amount which, together with the administered IFN-α

, is an effective IFN-α

treatment, wherein the amount of administered IFN-α

is less than an amount of IFN-α

required in the absence of coadministering the immunostimulatory nucleic acid, and wherein said immunostimulatory nucleic acid is 10 to 100 nucleotides long and comprises a poly-G sequence at each end and a central palindromic sequence comprising an unmethylated CpG dinucleotide.

52. The method of claim 19, wherein the coadministering comprises administering the IFN-α

and the immunostimulatory nucleic acid together.

53. The method of claim 19, wherein the coadministering comprises administering the IFN-α

and the immunostimulatory nucleic acid sequentially.

54. The method of claim 19, wherein the amount of administered IFN-α

is at least 20 percent below an amount of IFN-α

required in absence of coadministering the immunostimulatory nucleic acid.

55. The method of claim 19, wherein the amount of administered IFN-α

is at least 30 percent below an amount of IFN-α

required in absence of coadministering the immunostimulatory nucleic acid.

56. The method of claim 19, wherein the amount of administered IFN-α

is at least 40 percent below an amount of IFN-α

required in absence of coadministering the immunostimulatory nucleic acid.

57. The method of claim 19, wherein the amount of administered IFN-α

is at least 50 percent below an amount of IFN-α

required in absence of coadministering the immunostimulatory nucleic acid.

58. The method of claim 19, wherein the pharmaceutical composition comprising an immunostimulatory nucleic acid is administered locally.

59. The method of claim 19, wherein the immunostimulatory nucleic acid is stabilized.

60. The method of claim 19, wherein the immunostimulatory nucleic acid comprises a backbone with at least one nuclease-resistant internucleotide linkage selected from the group consisting of:

phosphorothioate, phosphorodithioate, methylphosphonate, and peptide.

61. The method of claim 19, wherein the immunostimulatory nucleic acid comprises at least one nucleotide analog or derivative.

62. The method of claim 19, wherein the immunostimulatory nucleic acid is between 12 and 40 nucleotides in length.

63. The method of claim 19, wherein the immunostimulatory nucleic acid has a sequence selected from the group consisting of

ggGGGACGATCGTCgggggGODN 2216SEQ ID NO:

64. The method of claim 19, wherein the subject has a condition selected from the group consisting of a proliferative disorder and a viral infection.

65. The method of claim 19, wherein the subject has a proliferative disorder selected from the group consisting of:

hairy cell leukemia, chronic myelogenous leukemia, cutaneous T-cell leukemia, multiple myeloma, follicular lymphoma, malignant melanoma, squamous cell carcinoma, AIDS-related Kaposi'"'"'s sarcoma, renal cell carcinoma, prostate carcinoma, bladder cell carcinoma, cervical dysplasia, and colon carcinoma.

66. The method of claim 19, wherein the subject has a viral infection selected from the group consisting of:

hepatitis B, hepatitis C, condyloma acuminatum, human immunodeficiency virus, herpes, cytomegalovirus, Epstein-Barr virus, and papillomavirus.

20. A method of reducing an interferon alpha (IFN-α

) treatment-related side effect in a subject receiving or in need of treatment with IFN-α

, comprising

administering to a mammalian subject in need of treatment with IFN-α

a pharmaceutical composition comprising IFN-α

, and

coadministering to the subject in need of such treatment a pharmaceutical composition comprising an immunostimulatory nucleic acid in an amount which, together with the administered IFN-α

c, is an effective IFN-α

treatment, wherein an IFN-α

treatment-related side effect is reduced in comparison to the side effect when IFN-α

is administered in the absence of coadministering the immunostimulatory nucleic acid, and wherein said immunostimulatory nucleic acid is 10 to 100 nucleotides long and comprises a poly-G sequence at each end and a central palindromic sequence comprising an unmethylated CpG dinucleotide.

67. The method of claim 20, wherein the co-administering comprises administering the IFN-α

and the immunostimulatory nucleic acid together.

68. The method of claim 20, wherein the co-administering comprises administering the IFN-α

and the immunostimulatory nucleic acid sequentially.

69. The method of claim 20, wherein the pharmaceutical composition comprising an immunostimulatory nucleic acid is administered locally.

70. The method of claim 20, wherein the IFN-α

treatment-related side effect is systemic.

71. The method of claim 20, wherein the IFN-α

treatment-related side effect is selected from the group consisting of flu-like syndrome, fever, headache, chills, myalgia, fatigue, anorexia, nausea, vomiting, diarrhea, and depression.

72. The method of claim 20, wherein the immunostimulatory nucleic acid is stabilized.

73. The method of claim 20, wherein the immunostimulatory nucleic acid comprises a backbone with at least one nuclease-resistant internucleotide linkage selected from the group consisting of:

phosphorothioate, phosphorodithioate, methylphosphonate, and peptide.

74. The method of claim 20, wherein the immunostimulatory nucleic acid comprises at least one nucleotide analog or derivative.

75. The method of claim 20, wherein the immunostimulatory nucleic acid is between 12 and 40 nucleotides in length.

76. The method of claim 20, wherein the immunostimulatory nucleic acid has a sequence selected from the group consisting of

ggGGGACGATCGTCgggggGODN 2216SEQ ID NO:

77. The method of claim 20, wherein the subject has a condition selected from the group consisting of a proliferative disorder and a viral infection.

78. The method of claim 20, wherein the subject has a proliferative disorder selected from the group consisting of:

hairy cell leukemia, chronic myelogenous leukemia, cutaneous T-cell leukemia, multiple myeloma, follicular lymphoma, malignant melanoma, squamous cell carcinoma, AIDS-related Kaposi'"'"'s sarcoma, renal cell carcinoma, prostate carcinoma, bladder cell carcinoma, cervical dysplasia, and colon carcinoma.

79. The method of claim 20, wherein the subject has a viral infection selected from the group consisting of:

hepatitis B, hepatitis C, condyloma acuminatum, human immunodeficiency virus, herpes, cytomegalovirus, Epstein-Barr virus, and papillomavirus.

21. A method of stimulating production of a plurality of type I interferon (IFN) subtypes in a mammalian subject, comprising administering to a mammalian subject in need of IFN-α

treatment an amount of immunostimulatory nucleic acid effective to induce IFN-producing cells (IPCs) to secrete at least two type I interferons, wherein said immunostimulatory nucleic acid is selected from the group consisting of

ggGGGACGATCGTCgggggGODN 2216SEQ ID NO;

7ggGGGACGATATCGTCgggggGODN 2245SEQ ID NO;

9ggGGGACGAGCTCGTCgggggGODN 2247SEQ ID NO;

11ggGGGACGATCGTTGggggGODN 2252SEQ ID NO;

13ggGGTCACCGGTGAgggggGODN 2300SEQ ID NO;

24ggGGTCGACGTACGTCGAgggggGODN 2301SEQ ID NO;

25ggGGACGTCGACGTggggGODN 2306SEQ ID NO;

30ggGTCGTCGACGAggggggGODN 2329SEQ ID NO;

33ggGGTCGACGTCGACGTC9AGgggggGODN 2334SEQ ID NO;

36, andggGGACGACGTCGTGgggggGODN 2336SEQ ID NO;

37,

wherein each lower case letter represents phosphorothioate linkage and each upper case letter indicates phosphodiester linkage.

80. The method of claim 21, wherein the IPCs are precursor type 2 dendritic cells (pDC2s).

81. The method of claim 21, wherein the IPCs are induced to secrete at least three type I interferons.

82. The method of claim 21, wherein the IPCs are induced to secrete at least four type I interferons.

83. The method of claim 21, wherein the IPCs are induced to secrete at least five type I interferons.

84. The method of claim 21, wherein the IPCs are induced to secrete at least six type I interferons.

85. The method of claim 21, wherein the IPCs are induced to secrete at least seven type I interferons.

86. The method of claim 21, wherein the IPCs are induced to secrete at least eight type I interferons.

87. The method of claim 21, wherein the immunostimulatory nucleic acid is stabilized.

88. The method of claim 21, wherein the immunostimulatory nucleic acid comprises a backbone with at least one nuclease-resistant internucleotide linkage selected from the group consisting of:

phosphorothioate, phosphorodithioate, methylphosphonate, and peptide.

89. The method of claim 21, wherein the immunostimulatory nucleic acid comprises at least one nucleotide analog or derivative.

22. A pharmaceutical composition, comprising

an isolated nucleic acid having a sequence selected from the group consisting of:

ggGGGACGATCGTCgggggGODN 2216SEQ ID NO;

7ggGGGACGATATCGTCgggggGODN 2245SEQ ID NO;

9ggGGGACGAGCTCGTCgggggGODN 2247SEQ ID NO;

11ggGGGACGATCGTTGggggGODN 2252SEQ ID NO;

13ggGGTCACCGGTGAgggggGODN 2300SEQ ID NO;

24ggGGTCGACGTACGTCGAgggggGODN 2301SEQ ID NO;

25ggGGACGTCGACGTggggGODN 2306SEQ ID NO;

30ggGTCGTCGACGAggggggGODN 2329SEQ ID NO;

33ggGGTCGACGTCGACGTCGAGgggggGODN 2334SEQ ID NO;

36, andggGGACGACGTCGTGgggggGODN 2336SEQ ID NO;

37,

wherein each lower case letter represents phosphorothioate linkage and each upper case letter indicates phosphodiester linkage; and

a pharmaceutically acceptable carrier.

23. The pharmaceutical composition of claim 22, further comprising IFN-α

.