Methods of inhibiting B lymphocytes using antibodies to Neutrokine-alpha

• US 8,071,092 B1
• Filed: 06/08/2000
• Issued: 12/06/2011
• Est. Priority Date: 10/25/1996
• Status: Expired due to Fees

• Current Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Original Assignee: Human Genome Sciences Incorporated (GlaxoSmithKline PLC)
• Inventors: Yu, Guo-Liang, Ni, Jian, Ebner, Reinhard
• Primary Examiner(s): Bunner, Bridget E
• Application Number:
  US09/589,288
• Time in Patent Office: 4,198 Days
• Field of Search: 514/2, 514/4, 514/8, 514/12, 530/300, 530/350, 530/387.1, 424/85.1, 424/130.1, 424/139.1, 424/13, 435/69.1, 435/69.5, 435/325, 435/328, 435/331, 435/335
• US Class Current: 424/130.1
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/52 Cytokines; Lymphokines; Interferons
• CPC Class Codes: C07K 14/715 for cytokines; for lymphokines; for interferons
• CPC Class Codes: C07K 16/24 against cytokines, lymphokines or interferons
• CPC Class Codes: C07K 16/2875 against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154 against NGF C07K16/22, against TNF C07K16/241
• CPC Class Codes: C07K 2317/34 Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/76 Antagonist effect on antigen, e.g. neutralization or inhibition of binding
• CPC Class Codes: G01N 2333/52 Assays involving cytokines
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 2500/02 Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
• CPC Class Codes: G01N 2800/245 : Transplantation related diseases, e.g. graft versus host disease
• CPC Class Codes: G01N 33/564 : for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
• CPC Class Codes: G01N 33/6863 : Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
• 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

The present invention relates to a novel Neutrokine-alpha, and a splice variant thereof designated Neutrokine-alphaSV, polynucleotides and polypeptides which are members of the TNF family. In particular, isolated nucleic acid molecules are provided encoding the human Neutrokine-alpha and/or Neutrokine-alphaSV polypeptides, including soluble forms of the extracellular domain. Neutrokine-alpha and/or Neutrokine-alphaSV polypeptides are also provided as are vectors, host cells and recombinant methods for producing the same. The invention further relates to screening methods for identifying agonists and antagonists of Neutrokine-alpha and/or Neutrokine-alphaSV activity. Also provided are diagnostic methods for detecting immune system-related disorders and therapeutic methods for treating immune system-related disorders.

Claims

1. A method of inhibiting B lymphocytes comprising administering an effective amount of an antibody that binds a protein whose amino acid sequence is

MDDSTEREQS RLTSCLKKRE EMKLKECVSI LPRKESPSVR SSKDGKLLAA TLLLALLSCC LTVVSFYQVA ALQGDLASLR AELQGHHAEK LPAGAGAPKA GLEEAPAVTA GLKIFEPPAP GEGNSSQNSR NKRAVQGPEE TVTQDCLQLI ADSETPTIQK GSYTFVPWLL SFKRGSALEE KENKILVKET GYFFIYGQVL YTDKTYAMGH LIQRKKVHVF GDELSLVTLF RCIQNMPETL PNNSCYSAGI AKLEEGDELQ LAIPRENAQI SLDGDVTFFG ALKLL (SEQ ID NO:

2)

wherein B lymphocytes are inhibited.

4. The method of any one of claims 1-3, wherein the antibody is a monoclonal antibody.

5. The method of any one of claims 1-3, wherein the antibody is recombinantly produced.

6. The method of any one of claims 1-3, wherein the antibody is a chimeric antibody.

7. The method of any one of claims 1-3, wherein the antibody is a humanized antibody.

8. The method of any one of claims 1-3, wherein the antibody comprises human constant domains.

9. The method of any one of claims 1-3, wherein the antibody is a F(ab′

)2 fragment.

10. The method of any one of claims 1-3, wherein the antibody is a polyclonal antibody.

11. The method of any one of claims 1-3, wherein the antibody is a Fab fragment.

12. The method of any one of claims 1-3, wherein the antibody is administered to an individual.

13. The method of any one of claims 1-3, wherein the antibody is administered to a cell culture.

2. A method of inhibiting B lymphocyte proliferation comprising administering an effective amount of an antibody that binds Neutrokine-alpha (SEQ ID NO:

2), wherein B lymphocyte proliferation is inhibited.

3. A method of inhibiting B lymphocyte differentiation comprising administering an effective amount of an antibody that binds Neutrokine-alpha (SEQ ID NO:

2), wherein B lymphocyte differentiation is inhibited.

14. A method of treating an autoimmune disease or disorder comprising administering to an individual, an effective amount of an antagonistic antibody or portion thereof that specifically binds a protein consisting of the amino acid sequence of amino acid residues 134-285 of SEQ ID NO:

2.

15. The method of claim 14 wherein the antibody or portion thereof is a monoclonal antibody.

16. The method of claim 14 wherein the antibody or portion thereof is a polyclonal antibody.

17. The method of claim 14 wherein the antibody or portion thereof is a Fab fragment.

18. The method of claim 14 wherein the antibody or portion thereof is labeled.

19. The method of claim 18 wherein the label is selected from the group consisting of:

(a) an enzyme label;

(b) a radioisotope;

(c) a fluorescent label; and

(d) biotin.

20. The method of claim 19 wherein the label is a radioisotope selected from the group consisting of:

(a) ¹²⁵I;

(b) ¹²¹I;

(c) ¹³¹I;

(d) ¹¹²In; and

(e) ^99mTc.

21. The method of claim 14 wherein the autoimmune disease or disorder is systemic lupus erythematosus.

22. The method of claim 14 wherein said antibody or portion thereof is administered intravenously.

23. The method of claim 14 which comprises administering between 0.1 and 20 mg/kg of the patient'"'"'s body weight of said antibody or portion thereof.

24. The method of claim 14, wherein the autoimmune disease or disorder is selected from the group consisting of systemic lupus erythematosus, idiopathic thrombocytopoietic purpura (ITP), Sjogren'"'"'s syndrome, Waldenstrom'"'"'s macroglobulinaemia, multiple sclerosis, cancer, asthma, nephritis, diabetes, scleroderma, vasculitis, cryoglobulinaemia, graft-versus-host disease (GVHD), renal transplantation, and antiphospholipid syndrome.

25. The method of claim 24, wherein the antibody or portion thereof is a monoclonal antibody.

26. The method of claim 24, wherein the antibody or portion thereof is a polyclonal antibody.

27. The method of claim 24, wherein the antibody or portion thereof is a Fab fragment.

28. The method of claim 24, wherein the antibody or portion thereof is labeled.

29. The method of claim 28, wherein the label is selected from the group consisting of:

(a) an enzyme label;

(b) a radioisotope;

(c) a fluorescent label; and

(d) biotin.

30. The method of claim 29, wherein the label is a radioisotope selected from the group consisting of:

(a) ¹²⁵I;

(b) ¹²¹I;

(c) ¹³¹I;

(d) ¹¹²In; and

(e) ^99mTc.

31. The method of claim 24, wherein the antibody or portion thereof is administered intravenously.

32. The method of claim 24, wherein the method comprises administering between 0.1 and 20 mg/kg of the patient'"'"'s body weight of the antibody or portion thereof.

33. The method of claim 24, wherein the autoimmune disease or disorder is idiopathic thrombocytopietic purpura (ITP).

34. The method of claim 24, wherein the autoimmune disease or disorder is Sjogren'"'"'s syndrome.

35. The method of claim 24, wherein the autoimmune disease or disorder is multiple sclerosis.

36. The method of claim 24, wherein the autoimmune disease or disorder is renal transplantation.

37. A method of treating rheumatoid arthritis comprising administering to an individual, an effective amount of an antagonistic antibody or portion thereof that specifically binds a protein consisting of the amino acid sequence of amino acid residues 134-285 of SEQ ID NO:

2.

38. The method of claim 37 wherein the antibody or portion thereof is a monoclonal antibody.

39. The method of claim 37 wherein the antibody or portion thereof is a polyclonal antibody.

40. The method of claim 37 wherein the antibody or portion thereof is a Fab fragment.

41. The method of claim 37 wherein the antibody or portion thereof is labeled.

42. The method of claim 41 wherein the label is selected from the group consisting of:

(a) an enzyme label;

(b) a radioisotope;

(c) a fluorescent label; and

(d) biotin.

43. The method of claim 42 wherein the label is a radioisotope selected from the group consisting of:

(a) ¹²⁵I;

(b) ¹²¹I;

(c) ¹³¹I;

(d) ¹¹²In; and

(e) ^99mTc.

44. The method of claim 37 wherein said antibody or portion thereof is administered intravenously.

45. The method of claim 37 which comprises administering between 0.1 and 20 mg/kg of the patient'"'"'s body weight of said antibody or portion thereof.

46. A method of inhibiting B lymphocyte proliferation, differentiation or survival comprising administering to an individual or a cell culture containing B lymphocytes, an effective amount of an antagonistic antibody or portion thereof that specifically binds a protein consisting of an amino acid sequence selected from the group consisting of:

(a) the amino acid sequence of amino acid residues n to 285 of SEQ ID NO;

2, where n is an integer in the range of 2-190;

(b) the amino acid sequence of amino acid residues 1 to m of SEQ ID NO;

2, where m is an integer in the range of 274 to 284; and

(c) the amino acid sequence of amino acid residues n to m of SEQ ID NO;

2, where n is an integer in the range of 2-190 and m is an integer in the range of 274-284.

47. The method of claim 46 wherein the protein consists of amino acid sequence (a).

48. The method of claim 46 wherein the protein consists of amino acid sequence (b).

49. The method of claim 46 wherein the protein consists of amino acid sequence (c).

50. The method of claim 46 wherein the antibody or portion thereof is a monoclonal antibody.

51. The method of claim 46 wherein the antibody or portion thereof is a polyclonal antibody.

52. The method of claim 46 wherein the antibody or portion thereof is a Fab fragment.

53. The method of claim 46 wherein the antibody or portion thereof is labeled.

54. The method of claim 53 wherein the label is selected from the group consisting of:

(a) an enzyme label;

(b) a radioisotope;

(c) a fluorescent label; and

(d) biotin.

55. The method of claim 54 wherein the label is a radioisotope selected from the group consisting of:

(a) ¹²⁵I;

(b) ¹²¹I;

(c) ¹³¹I;

(d) ¹¹²In; and

(e) ^99mTc.

56. The method of claim 46 which comprises administering to an individual an effective amount of said antagonistic antibody or portion thereof.

57. The method of claim 46 which comprises administering to a cell culture containing B lymphocytes an effective amount of said antagonistic antibody or portion thereof.

58. A method of inhibiting B lymphocyte proliferation, differentiation, or survival comprising administering to an individual or a cell culture containing B lymphocytes, an effective amount of an antagonistic antibody or portion thereof that specifically binds a protein consisting of the amino acid sequence of amino acid residues 134-285 of SEQ ID NO:

2.

59. The method of claim 58 wherein the antibody or portion thereof is a monoclonal antibody.

60. The method of claim 58 wherein the antibody or portion thereof is a polyclonal antibody.

61. The method of claim 58 wherein the antibody or portion thereof is a Fab fragment.

62. The method of claim 58 wherein the antibody or portion thereof is labeled.

63. The method of claim 62 wherein the label is selected from the group consisting of:

(a) an enzyme label;

(b) a radioisotope;

(c) a fluorescent label; and

(d) biotin.

64. The method of claim 63 wherein the label is a radioisotope selected from the group consisting of:

(a) ¹²⁵I;

(b) ¹²¹I;

(c) ¹³¹I;

(d) ¹¹²In; and

(e) ^99mTc.

65. The method of claim 58 which comprises administering to an individual an effective amount of said antagonistic antibody or portion thereof.

66. The method of claim 58 which comprises administering to a cell culture containing B lymphocytes an effective amount of said antagonistic antibody or portion thereof.

67. A method of treating an autoimmune disease or disorder comprising administering to an individual, an effective amount of an antagonistic antibody or portion thereof that specifically binds to an isolated recombinant Neutrokine-a protein purified from a cell culture wherein the cells in said cell culture comprise a polynucleotide encoding amino acids 1-285 of SEQ ID NO:

2 operably associated with a regulatory sequence that controls gene expression.

68. The method of claim 67 wherein the antibody or portion thereof is a monoclonal antibody.

69. The method of claim 67 wherein the antibody or portion thereof is a polyclonal antibody.

70. The method of claim 67 wherein the antibody or portion thereof is a Fab fragment.

71. The method of claim 67 wherein the antibody or portion thereof is labeled.

72. The method of claim 71 wherein the label is selected from the group consisting of:

(a) an enzyme label;

(b) a radioisotope;

(c) a fluorescent label; and

(d) biotin.

73. The method of claim 72 wherein the label is a radioisotope selected from the group consisting of:

(a) ¹²⁵I;

(b) ¹²¹I;

(c) ¹³¹I;

(d) ¹¹²In; and

(e) ^99mTc.

74. The method of claim 67 wherein the autoimmune disease or disorder is systemic lupus erythematosus.

75. The method of claim 67 wherein said antibody or portion thereof is administered intravenously.

76. The method of claim 67 which comprises administering between 0.1 and 20 mg/kg of the patient'"'"'s body weight of said antibody or portion thereof.

77. The method of claim 67, wherein the autoimmune disease or disorder is selected from the group consisting of systemic lupus erythematosus, idiopathic thrombocytopoietic purpura (ITP), Sjogren'"'"'s syndrome, Waldenstrom'"'"'s macroglobulinaemia, multiple sclerosis, cancer, asthma, nephritis, diabetes, scleroderma, vasculitis, cryoglobulinaemia, graft-versus-host disease (GVHD), renal transplantation, and antiphospholipid syndrome.

78. The method of claim 77, wherein the autoimmune disease or disorder is systemic lupus erythematosus.

79. The method of claim 77, wherein the autoimmune disease or disorder is idiopathic thrombocytopietic purpura (ITP).

80. The method of claim 77, wherein the autoimmune disease or disorder is Sjogren'"'"'s syndrome.

81. The method of claim 77, wherein the autoimmune disease or disorder is multiple sclerosis.

82. The method of claim 77, wherein the autoimmune disease or disorder is renal transplantation.

83. A method of treating rheumatoid arthritis comprising administering to an individual, an effective amount of an antagonistic antibody or portion thereof that specifically binds to an isolated recombinant Neutrokine-a protein purified from a cell culture wherein the cells in said cell culture comprise a polynucleotide encoding amino acids 1-285 of SEQ ID NO:

2 operably associated with a regulatory sequence that controls gene expression.

84. The method of claim 83 wherein the antibody or portion thereof is a monoclonal antibody.

85. The method of claim 83 wherein the antibody or portion thereof is a polyclonal antibody.

86. The method of claim 83 wherein the antibody or portion thereof is a Fab fragment.

87. The method of claim 83 wherein the antibody or portion thereof is labeled.

88. The method of claim 87 wherein the label is selected from the group consisting of:

(a) an enzyme label;

(b) a radioisotope;

(c) a fluorescent label; and

(d) biotin.

89. The method of claim 88 wherein the label is a radioisotope selected from the group consisting of:

(a) ¹²⁵I;

(b) ¹²¹I;

(c) ¹³¹I;

(d) ¹¹²In; and

(e) ^99mTc.

90. The method of claim 83 wherein said antibody or portion thereof is administered intravenously.

91. The method of claim 83 which comprises administering between 0.1 and 20 mg/kg of the patient'"'"'s body weight of said antibody or portion thereof.

92. A method of treating an autoimmune disease in an animal comprising administering a therapeutically effective amount of an anti-Neutrokine-alpha antibody that binds to human Neutrokine alpha polypeptide having the amino acid sequence of SEQ ID NO:

2.

93. The method of claim 92, wherein the antibody is a monoclonal antibody.

94. The method of claim 92, wherein the antibody is recombinantly produced.

95. The method of claim 92, wherein the antibody is a chimeric antibody.

96. The method of claim 92, wherein the antibody is a humanized antibody.

97. The method of claim 92, wherein the antibody comprises human constant domains.

98. The method of claim 92, wherein the antibody is a F(ab′

)2 fragment.

99. The method of claim 92, wherein the animal is human.

100. The method of claim 92, further comprising detecting the levels of B cell growth and immunoglobulin production in the animal.

101. The method of claim 92, further comprising detecting the level of B cell growth in the animal.

102. The method of claim 92, further comprising detecting the level of immunoglobulin production in the animal.