Fusion proteins for blood-brain barrier delivery

• US 20070081992A1
• Filed: 10/07/2005
• Published: 04/12/2007
• Est. Priority Date: 10/07/2005
• Status: Active Grant

• Current Assignee: JCR Pharmaceuticals Co., Ltd.
• Original Assignee: ArmaGen Technologies, Inc.
• Inventors: Pardridge, William, Boado, Ruben
• Granted Patent: US 8,142,781 B2
• Time in Patent Office: Days
• US Class Current: 424/143.100
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/24 Antidepressants
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C07K 14/475 Growth factors; Growth regulators
• CPC Class Codes: C07K 16/2869 against hormone receptors for antibodies against neuromediator receptors C07K16/286
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

Abstract

The invention provides compositions, methods, and kits for increasing transport of agents across the blood brain barrier while allowing their activity once across the barrier to remain substantially intact. The agents are transported across the blood brain barrier via one or more endogenous receptor-mediated transport systems. In some embodiments the agents are therapeutic, diagnostic, or research agents.

Claims

1. A composition comprising a neurotherapeutic agent covalently linked to a structure that is capable of crossing the blood brain barrier (BBB), wherein the composition is capable of producing an average elevation of concentration in the brain of the neurotherapeutic agent of at least about 5 ng/gram brain following peripheral administration.

2. The composition of claim 1 wherein the neurotherapeutic agent has a molecular weight greater than about 400 Daltons.

3. The composition of claim 1 wherein the neurotherapeutic agent alone does not cross the BBB in a therapeutically effective amount following peripheral administration.

4. The composition of claim 1 wherein the structure that is capable of crossing the BBB crosses the BBB on an endogenous BBB receptor mediated transport system.

5. The composition of claim 4 wherein the endogenous BBB receptor mediated transport system is selected from the group consisting of the insulin receptor, transferrin receptor, leptin receptor, lipoprotein receptor, and the IGF receptor.

6. The composition of claim 5 wherein the endogenous BBB receptor mediated transport system is the insulin BBB receptor mediated transport system.

7. The composition of claim 1 wherein the structure that is capable of crossing the BBB is an antibody.

8. The composition of claim 7 wherein the antibody is a monoclonal antibody (MAb).

9. The composition of claim 8 wherein the MAb is a chimeric MAb.

10. The composition of claim 9 wherein the chimeric antibody contains sufficient human sequences to avoid significant immunogenic reaction when administered to a human.

11. The composition of claim 5 wherein the structure that crosses the BBB on an endogenous BBB receptor mediated transport system is an antibody.

12. The composition of claim 11 wherein the antibody is a monoclonal antibody.

13. The composition of claim 12 wherein the MAb is a chimeric MAb.

14. The composition of claim 13 wherein the chimeric antibody contains sufficient human sequences to avoid significant immunogenic reaction when administered to a human.

15. The composition of claim 1 wherein the neurotherapeutic agent is a neurotrophin.

16. The composition of claim 15 wherein the neurotrophin is selected from the group consisting of brain derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurotrophin-4/5, fibroblast growth factor (FGF)-2 and other FGFs, neurotrophin (NT)-3, erythropoietin (EPO), hepatocyte growth factor (HGF), epidermal growth factor (EGF), transforming growth factor (TGF)-α

, TGF-β

, vascular endothelial growth factor (VEGF), interleukin-1 receptor antagonist (IL-1ra), ciliary neurotrophic factor (CNTF), glial-derived neurotrophic factor (GDNF), neurturin, platelet-derived growth factor (PDGF), heregulin, neuregulin, artemin, persephin, interleukins, granulocyte-colony stimulating factor (CSF), granulocyte-macrophage-CSF, netrins, cardiotrophin-1, hedgehogs, leukemia inhibitory factor (LIF), midkine, pleiotrophin, bone morphogenetic proteins (BMPs), netrins, saposins, semaphorins, and stem cell factor (SCF).

17. The composition of claim 15 wherein the neurotrophin comprises brain derived neurotrophic factor (BDNF).

18. The composition of claim 17 wherein the BDNF is a variant of native BDNF.

19. The composition of claim 17 wherein the BDNF is a two amino acid carboxyl-truncated variant.

20. The composition of claim 17 or 19 wherein the BDNF is a human BDNF.

21. The composition of claim 19 wherein the BDNF comprises a sequence that is at least about 80% identical to the sequence of amino acids 466-582 of SEQ ID NO:

24.

22. The composition of claim 1 wherein the neurotherapeutic agent is a neurotrophin and the structure that is capable of crossing the blood brain barrier (BBB) is a MAb to an endogenous BBB receptor mediated transport system.

23. The composition of claim 22 wherein the neurotrophin is BDNF.

24. The composition of claim 23 wherein the MAb is an antibody to the insulin BBB receptor mediated transport system.

25. The composition of claim 24 wherein the BDNF is a two amino acid carboxy-truncated BDNF.

26. The composition of claim 25 wherein the MAb is a chimeric MAb

27. The composition of claim 26 wherein the chimeric antibody contains sufficient human sequences to avoid significant immunogenic reaction when administered to a human.

28. The composition of claim 26 wherein the insulin receptor is a human insulin receptor and wherein the BDNF is a human BDNF.

29. The composition of claim 28 wherein the BDNF comprises a sequence that is at least about 80% identical to the sequence of amino acids 466-582 of SEQ ID NO:

24.

30. The composition of claim 29 wherein the BDNF is covalently linked at its amino terminus to the carboxy terminus of the heavy chain of the MAb.

31. The compostion of claim 29 wherein the BDNF is covalently linked at its amino terminus to the carboxy terminus of the light chain of the MAb.

32. The composition of claim 30 wherein the heavy chain of the MAb comprises a sequence that is at least about 80% identical to amino acids 20-462 of SEQ ID NO:

24.

33. The composition of claim 32 further comprising a linker between the heavy chain of the MAb and the BDNF.

34. The composition of claim 33 wherein the linker is S-S-M.

35. The composition of claim 34 further comprising the light chain of the MAb.

36. The composition of claim 35 wherein the light chain comprises a sequence that is at least about 80% identical to amino acids 21-234 of SEQ ID NO:

36.

37. The composition of claim 36 wherein the MAb is glycosylated.

38. A pharmaceutical composition comprising the composition of claim 1, 18, 21, 30, or 31 and a pharmaceutically acceptable excipient.

39. A composition comprising the composition of claim 1 and second composition comprising a second neurotherapeutic agent covalently linked to a second structure that is capable of crossing the blood brain barrier (BBB).

40. The composition of claim 39 wherein the first and second neurotherapeutic agents are different and the first and second structures capable of crossing the BBB are the same structure.

41. The composition of claim 40 wherein the structure capable of crossing the BBB is an antibody comprising a first heavy chain and a second heavy chain.

42. The composition of claim 40 wherein the first neurotherapeutic agent is covalently linked to the first heavy chain of the antibody and the second neurotherapeutic agent is covalently linked to the second heavy chain of the antibody.

43. A composition comprising an agent covalently linked to a chimeric MAb to the human BBB insulin receptor, wherein the MAb comprises a heavy chain and a light chain.

44. The composition of claim 43 wherein the agent is a therapeutic agent.

45. The composition of claim 44 wherein the therapeutic agent is a neurotrophin.

46. The composition of claim 45 wherein the neurotrophin is a BDNF.

47. The composition of claim 46 wherein the agent is a two amino acid carboxyl-terminal truncated BDNF.

48. The composition of claim 46 wherein the heavy chain of the MAb is covalently linked to the BDNF to form a fusion protein, and wherein the sequence of said fusion protein comprises a first sequence that is at least about 80% identical to a sequence comprising amino acids 20-462 of SEQ ID NO:

24 and further comprises a second sequence that is at least about 80% identical to a sequence comprising amino acids 466-582 of SEQ ID NO;

24.

49. The composition of claim 48 wherein the light chain of the MAb comprises a sequence that is at least about 80% identical to a sequence comprising amino acids 21-234 of SEQ ID NO:

36.

50. The composition of claim 48 wherein the MAb is glycosylated.

51. The composition of claim 49 further comprising a peptide linker between the carboxyl terminus of the first sequence and the amino terminus of the second sequence.

52. The composition of claim 51 wherein the linker comprises S-S-M.

53. The composition of claim 52 wherein the MAb is glycosylated.

54. A composition for treating a neurological disorder comprising a BDNF covalently linked to an immunoglobulin that is capable of crossing the blood brain barrier, wherein the composition is capable of crossing the BBB in an amount that is effective in treating the neurological disorder.

55. A fusion protein comprising:

(i) a structure capable of crossing the BBB, covalently linked to

(ii) a peptide that is active in the central nervous system (CNS),

wherein the structure capable of crossing the blood brain barrier and the peptide that is active in the central nervous system each retain an average of at least about 40% of their. activities, compared to their activities as separate entities.

56. The fusion protein of claim 55 wherein the structure capable of crossing the blood brain barrier crosses the BBB on an endogenous BBB receptor-mediated transporter.

57. The fusion protein of claim 56 wherein the endogenous BBB receptor mediated transport system is selected from the group consisting of the insulin receptor, transferrin receptor, leptin receptor, lipoprotein receptor, and the IGF receptor.

58. The fusion protein of claim 57 wherein the endogenous BBB receptor-mediated transporter is selected from the group consisting of the insulin transporter and the transferrin transporter.

59. The fusion protein of claim 58 wherein the endogenous BBB receptor-mediated transporter is the insulin transporter.

60. The fusion protein of claim 59 wherein the insulin transporter is the human insulin transporter.

61. The fusion protein of claim 55 wherein the structure capable of crossing the BBB is an antibody.

62. The fusion protein of claim 61 wherein the antibody is a MAb.

63. The fusion protein of claim 62 wherein the MAb is a chimeric MAb.

64. The fusion protein of claim 61 wherein the antibody is an antibody to an endogenous BBB receptor-mediated transporter

65. The fusion protein of claim 64 wherein the the endogenous BBB receptor mediated transport system is selected from the group consisting of the insulin receptor, transferrin receptor, leptin receptor, lipoprotein receptor, and the IGF receptor.

66. The fusion protein of claim 65 wherein the endogenous BBB receptor-mediated transporter is selected from the group consisting of the insulin transporter and the transferrin transporter.

67. The fusion protein of claim 66 wherein the endogenous BBB receptor-mediated transporter is the insulin transporter.

68. The fusion protein of claim 67 wherein the insulin transporter is the human insulin transporter.

69. The fusion protein of claim 55 wherein the peptide that is active in the CNS is a neurotherapeutic agent.

70. The fusion protein of claim 69 wherein the neurotherapeutic agent is a neurotrophin.

71. The fusion protein of claim 70 wherein the neurotrophin is selected from the group consisting of brain derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurotrophin-4/5, fibroblast growth factor (FGF)-2 and other FGFs, neurotrophin (NT)-3, erytropoietin (EPO), hepatocyte growth factor (HGF), epidermal growth factor (EGF), transforming growth factor (TGF)-α

, TGF-β

, vascular endothelial growth factor (VEGF), interleukin-1 receptor antagonist (IL-1ra), ciliary neurotrophic factor (CNTF), glial-derived neurotrophic factor (GDNF), neurturin, platelet-derived growth factor (PDGF), heregulin, neuregulin, artemin, persephin, interleukins, granulocyte-colony stimulating factor (CSF), granulocyte-macrophage-CSF, netrins, cardiotrophin-1, hedgehogs, leukemia inhibitory factor (LIF), midkine, pleiotrophin, bone morphogenetic proteins (BMPs), netrins, saposins, semaphorins, and stem cell factor (SCF).

72. The fusion protein of claim 71 wherein the neurotrophin is BDNF.

73. The fusion protein of claim 72 wherein the BDNF is a truncated BDNF.

74. The fusion protein of claim 73 wherein the truncated BDNF is a carboxyl-truncated BDNF.

75. The fusion protein of claim 74 wherein the carboxyl-truncated BDNF is lacking the two carboxyl terminal amino acids.

76. The fusion protein of claim 55 wherein the structure capable of crossing the BBB and the neurotherapeutic agent are covalently linked by a peptide linker.

77. A method of transport of an agent active in the CNS from the peripheral circulation across the BBB in an effective amount, comprising peripherally administering to an individual the agent covalently attached to a structure that crosses the BBB, under conditions where the agent covalently attached to a structure that crosses the BBB is transported across the BBB in an effective amount.

78. The method of claim 77 wherein the agent is a neurotherapeutic agent.

79. A method for treating a CNS disorder in an individual comprising peripherally administering to the individual an effective amount of a composition comprising a neurotherapeutic agent covalently attached to a structure capable of crossing the BBB.

80. The method of claim 79 wherein the structure capable of crossing the BBB comprises an antibody to an insulin receptor and the therapeutic agent comprises a BDNF.

81. The method of claim 80 wherein the administering is selected from the group consisting of oral, intravenous, intramuscular, subcutaneous, intraperitoneal, rectal, transbuccal, intranasal, transdermal, and inhalation administering

82. The method of claim 81 wherein the administering is intravenous, intramuscular, or subcutaneous.

83. The method of claim 79 wherein the CNS disorder is an acute CNS disorder.

84. The method of claim 83 wherein the acute CNS disorder is selected from the group consisting of spinal cord injury, brain injury focal brain ischemia and global brain ischemia.

85. The method of claim 83 wherein the composition is administered only once.

86. The method of claim 83 wherein the composition is administered at a frequency of no greater than about once per week.

87. The method of claim 79 wherein the CNS disorder is a chronic disorder.

88. The method of claim 87 wherein the chronic disorder is selected from the group consisting of chronic neurodegenerative disease, retinal ischemia, and depression.

89. The method of claim 88 wherein the chronic neurodegenerative disease is selected from the group consisting of prion diseases, Alzheimer'"'"'s disease, Parkinson'"'"'s disease, amyotrophic lateral sclerosis, Huntington'"'"'s disease, multiple sclerosis, transverse myelitis, motor neuron disease, Pick'"'"'s disease, tuberous sclerosis, lysosomal storage disorders, Canavan'"'"'s disease, Rett'"'"'s syndrome, spinocerebellar ataxias, Friedreich'"'"'s ataxia, optic atrophy, and retinal degeneration.

90. The method of claim 79 wherein the individual is a human.

91. The method of claim 90 wherein the individual is administered a dose of the composition that is about 1 to about 100 mg.

92. A composition comprising a cationic therapeutic peptide covalently linked to an immunoglobulin, wherein the cationic therapeutic peptide in the composition has a serum half-life that is an average of at least about 5-fold greater than the serum half-life of the cationic therapeutic peptide alone.

93. The composition of claim 92 wherein the cationic therapeutic peptide comprises a neurotherapeutic agent.

94. The composition of claim 93 wherein the neurotherapeutic agent is a neurotrophin.

95. The composition of claim 94 wherein the neurotrophin is selected from the group consisting of brain derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurotrophin-4/5, fibroblast growth factor (FGF)-2 and other FGFs, neurotrophin (NT)-3, erythropoietin (EPO), hepatocyte growth factor (HGF), epidermal growth factor (EGF), transforming growth factor (TGF)-α

, TGF-β

, vascular endothelial growth factor (VEGF), interleukin-1 receptor antagonist (IL-1ra), ciliary neurotrophic factor (CNTF), glial-derived neurotrophic factor (GDNF), neurturin, platelet-derived growth factor (PDGF), heregulin, neuregulin, artemin, persephin, interleukins, granulocyte-colony stimulating factor (CSF), granulocyte-macrophage-CSF, netrins, cardiotrophin-1, hedgehogs, leukemia inhibitory factor (LIF), midkine, pleiotrophin, bone morphogenetic proteins (BMPs), netrins, saposins, semaphorins, and stem cell factor (SCF).

96. The composition of claim 95 wherein the neurotrophin is BDNF.

97. The composition of claim 92 wherein the immunoglobulin is an antibody to an endogenous BBB receptor-mediated transport system.