Neuroprotective synergy of erythropoietin and insulin-like growth factors
View Patent ↗The present invention provides a method of providing acute neuroprotection by inducing the erythropoietin (EPO) signaling pathway in neuronal cells close to or subsequent to the time of excitatory insult; and inducing an insulin-like growth factor (IGF) signaling pathway in the neuronal cells close to or subsequent to the time of excitatory insult, thereby producing a synergistic acute neuroprotective effect in the neuronal cells. The invention also provides a method of preventing or reducing the severity of a neurologic condition in a subject by administering to the subject EPO or an active fragment or analog thereof at a dose of at most 2000 U/kg; and administering to the subject an IGF or an active fragment or analog thereof, thereby providing neuroprotection and preventing or reducing the severity of the neurologic condition. Such a method can be used to prevent or reduce the severity of, for example, Alzheimer's disease, Parkinson's disease, Huntington's disease, epilepsy, amyotrophic lateral sclerosis, multiple sclerosis, a movement disorder, HIV-associated dementia, HIV-associated neuropathy, neuropathic pain, migraine, glaucoma, drug addiction, drug withdrawal, drug dependency, depression or anxiety.
1. A method of providing neuroprotection from N-methyl-D-aspartic acid (NMDA) receptor-mediated excitatory insult, comprising:
(a) contacting neuronal cells in vitro with erythropoietin (EPO) close to or subsequent to the time of excitatory insult; and
(b) contacting said neuronal cells with an insulin-like growth factor (IGF) close to or subsequent to the time of excitatory insult,
thereby producing a synergistic neuroprotective effect from NMDA receptor-mediated excitatory insult in said neuronal cells.
2. The method of claim 1 , wherein said EPO is human EPO.
3. The method of claim 1 , wherein said EPO has at least 10-fold higher affinity for the EPO receptor than native human EPO.
4. The method of claim 1 , wherein said EPO is oligomeric.
5. The method of claim 4 , wherein said oligomeric EPO is dimeric.
6. The method of claim 1 , wherein said EPO has a half-life greater than native human EPO.
7. The method of claim 1 , wherein said EPO is hyper-glycosylated compared to native human EPO.
8. The method of claim 1 , further comprising contacting said neuronal cells with soluble EPO receptor.
9. The method of claim 1 , wherein said IGF is IGF-I.
10. The method of claim 9 , wherein said IGF-I is human IGF-I.
11. The method of claim 1 , wherein said IGF has at least 10-fold higher affinity for the IGF-I receptor than native human IGF-I.
12. The method of claim 1 , wherein said IGF active fragment has an altered affinity for an IGF-binding protein (IBP).
13. The method of claim 1 , wherein said IGF has a half-life greater than native human IGF.
14. A method of providing neuroprotection from NMDA receptor-mediated excitatory insult, comprising:
(a) contacting neuronal cells in vitro with an active fragment of erythropoietin (EPO) close to or subsequent to the time of excitatory insult; and
(b) contacting said neuronal cells with an insulin-like growth factor (IGF) close to or subsequent to the time of excitatory insult,
thereby producing a synergistic neuroprotective effect from NMDA receptor-mediated excitatory insult in said neuronal cells.
15. The method of claim 14 , wherein said EPO is human EPO.
16. The method of claim 14 , wherein said EPO active fragment has at least 10-fold higher affinity for the EPO receptor than native human EPO.
17. The method of claim 14 , wherein said EPO active fragment is oligomeric.
18. The method of claim 17 , wherein said oligomeric EPO active fragment is dimeric.
19. The method of claim 14 , wherein said EPO active fragment has a half-life greater than native human EPO.
20. The method of claim 14 , wherein said EPO active fragment is hyper-glycosylated compared to native human EPO.
21. The method of claim 14 , further comprising contacting said neuronal cells with soluble EPO receptor.
22. The method of claim 14 , wherein said IGF is IGF-I.
23. The method of claim 22 , wherein said IGF-I is human IGF-I.
24. The method of claim 14 , wherein said IGF has at least 10-fold higher affinity for the IGF-I receptor than native human IGF-I.
25. The method of claim 14 , wherein said IGF active fragment has an altered affinity for an IGF-binding protein (IBP).
26. The method of claim 14 , wherein said IGF has a half-life greater than native human IGF.
27. A method of providing neuroprotection from NMDA receptor-mediated excitatory insult, comprising:
(a) contacting neuronal cells in vitro with erythropoietin (EPO) close to or subsequent to the time of excitatory insult; and
(b) contacting said neuronal cells with an active fragment of insulin-like growth factor (IGF) close to or subsequent to the time of excitatory insult,
thereby producing a synergistic neuroprotective effect from NMDA receptor-mediated excitatory insult in said neuronal cells.
28. The method of claim 27 , wherein said EPO is human EPO.
29. The method of claim 27 , wherein said EPO has at least 10-fold higher affinity for the EPO receptor than native human EPO.
30. The method of claim 27 , wherein said EPO is oligomeric.
31. The method of claim 30 , wherein said oligomeric EPO is dimeric.
32. The method of claim 27 , wherein said EPO has a half life greater than native human EPO.
33. The method of claim 27 , wherein said EPO is hyper-glycosylated compared to native human EPO.
34. The method of claim 27 , further comprising contacting said neuronal cells with soluble EPO receptor.
35. The method of claim 27 , wherein said IGF active fragment is an IGF-I active fragment.
36. The method of claim 35 , wherein said IGF-I active fragment is an active fragment of human IGF-I.
37. The method of claim 27 , wherein said IGF active fragment has at least 10-fold higher affinity for the IGF-I receptor than native human IGF-I.
38. The method of claim 27 , wherein said IGF active fragment has an altered affinity for an IGF-binding protein (IBP).
39. The method of claim 27 , wherein said IGF active fragment has a half-life greater than native human IGF.
40. A method of providing neuroprotection from NMDA receptor-mediated excitatory insult, comprising:
(a) contacting neuronal cells in vitro with an active fragment of erythropoietin (EPO) close to or subsequent to the time of excitatory insult; and
(b) contacting said neuronal cells with an active fragment of insulin-like growth factor (IGF) close to or subsequent to the time of excitatory insult,
thereby producing a synergistic neuroprotective effect from NMDA receptor-mediated excitatory insult in said neuronal cells.
41. The method of claim 40 , wherein said EPO active fragment is an active fragment of human EPO.
42. The method of claim 40 , wherein said EPO active fragment has at least 10-fold higher affinity for the EPO receptor than native human EPO.
43. The method of claim 40 , wherein said EPO active fragment is oligomeric.
44. The method of claim 43 , wherein said oligomeric EPO active fragment is dimeric.
45. The method of claim 40 , wherein said EPO active fragment has a half-life greater than native human EPO.
46. The method of claim 40 , wherein said EPO active fragment is hyper-glycosylated compared to native human EPO.
47. The method of claim 40 , further comprising contacting said neuronal cells with soluble EPO receptor.
48. The method of claim 40 , wherein said IGF active fragment is an active fragment of IGF-I.
49. The method of claim 48 , wherein said IGF-I active fragment is an active fragment of human IGF-I.
50. The method of claim 40 , wherein said IGF active fragment has at least 10-fold higher affinity for the IGF-I receptor than native human IGF-I.
51. The method of claim 40 , wherein said IGF active fragment has an altered affinity for an IGF-binding protein (IBP).
52. The method of claim 40 , wherein said IGF active fragment has a half-life greater than native human IGF.
53. A method of providing neuroprotection from NMDA receptor-mediated excitatory insult, comprising:
(a) contacting neuronal cells in vitro with erythropoietin (EPO) close to or subsequent to the time of excitatory insult; and
(b) contacting said neuronal cells with an insulin-like growth factor (IGF) peptide analog close to or subsequent to the time of excitatory insult, wherein said IGF peptide analog is selected from SEQ ID NOS:13-20,
thereby producing a synergistic neuroprotective effect from NMDA receptor-mediated excitatory insult in said neuronal cells.
54. The method of claim 53 , wherein said EPO is human EPO.
55. The method of claim 53 , wherein said EPO has at least 10-fold higher affinity for the EPO receptor than native human EPO.
56. The method of claim 53 , wherein said EPO is oligomeric.
57. The method of claim 56 , wherein said oligomeric EPO is dimeric.
58. The method of claim 53 , wherein said EPO has a half-life greater than native human EPO.
59. The method of claim 53 , wherein said EPO is hyper-glycosylated compared to native human EPO.
60. The method of claim 53 , further comprising contacting said neuronal cells with soluble EPO receptor.
61. A method of providing neuroprotection from NMDA receptor-mediated excitatory insult, comprising:
(a) contacting neuronal cells in vitro with an active fragment of erythropoietin (EPO) close to or subsequent to the time of excitatory insult; and
(b) contacting said neuronal cells with an insulin-like growth factor (IGF) peptide analog close to or subsequent to the time of excitatory insult, wherein said IGF peptide analog is selected from SEQ ID NOS:13-20,
thereby producing a synergistic neuroprotective effect from NMDA receptor-mediated excitatory insult in said neuronal cells.
62. The method of claim 61 , wherein said EPO active fragment is an active fragment of human EPO.
63. The method of claim 61 , wherein said EPO active fragment has at least 10-fold higher affinity for the EPO receptor than native human EPO.
64. The method of claim 61 , wherein said EPO active fragment is oligomeric.
65. The method of claim 64 , wherein said oligomeric EPO active fragment is dimeric.
66. The method of claim 61 , wherein said EPO active fragment has a half-life greater than native human EPO.
67. The method of claim 61 , wherein said EPO active fragment is hyper-glycosylated compared to native human EPO.
68. The method of claim 61 , further comprising contacting said neuronal cells with soluble EPO receptor.