NEUROPROTECTIVE SYNERGY OF ERYTHROPOIETIN AND INSULIN-LIKE GROWTH FACTORS
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 acute neuroprotection, comprising:
(a) contacting neuronal cells with erythropoietin (EPO) or an active fragment or analog thereof close to or subsequent to the time of excitatory insult; and
(b) contacting said neuronal cells with an insulin-like growth factor (IGF) or an active fragment or analog thereof close to or subsequent to the time of excitatory insult,
thereby producing a synergistic acute neuroprotective effect in said neuronal cells.
2 . The method of claim 1 , wherein step (a) comprises contacting said neuronal cells with EPO or an active fragment thereof.
3 . The method of claim 2 , wherein said EPO is human EPO or an active fragment thereof.
4 . The method of claim 3 , comprising contacting said neuronal cells with human EPO.
5 . The method of claim 1 , wherein step (a) comprises contacting said neuronal cells with an EPO analog.
6 . The method of claim 5 , wherein said EPO analog is selected from the group consisting of GGTYSCHFGPLTWVCKPQGG (SEQ ID NO: 7);
GGDYHCRMGPLTWVCKPLGG (SEQ ID NO: 8);
GGVYACRMGPITWVCSPLGG (SEQ ID NO: 9);
VGNYMCHFGPITWVCRPGGG (SEQ ID NO: 10);
GGLYLCRFGPVTWDCGYKGG (SEQ ID NO: 11); and
GGCRIGPITWVCGG (SEQ ID NO: 12).
7 . The method of claim 5 , wherein said EPO analog is a peptidomimetic.
8 . The method of claim 5 , wherein said EPO analog is a small molecule.
9 . The method of claim 1 , wherein said EPO or active fragment or analog thereof has at least 10-fold higher affinity for the EPO receptor than native human EPO.
10 . The method of claim 1 , wherein said EPO or active fragment or analog thereof is oligomeric.
11 . The method of claim 10 , wherein said oligomeric EPO or active fragment or analog thereof is dimeric.
12 . The method of claim 11 , wherein said dimeric EPO or active fragment or analog thereof comprises dimeric GGTYSCHFGPLTWVCKPQGG (EMP1) (SEQ ID NO: 7).
13 . The method of claim 1 , wherein said EPO or active fragment or analog thereof has a half-life greater than native human EPO.
14 . The method of claim 1 , wherein said EPO or active fragment or analog thereof is hyper-glycosylated compared to native human EPO.
15 . The method of claim 14 , wherein step (a) comprises contacting said neuronal cells with Darbepoietin.
16 . The method of claim 1 , further comprising contacting said neuronal cells with soluble EPO receptor.
17 . The method of claim 1 , wherein step (b) comprises contacting said neuronal cells with an IGF or an active fragment thereof.
18 . The method of claim 17 , wherein said IGF or active fragment thereof is IGF-I or an active fragment thereof.
19 . The method of claim 18 , wherein said IGF-I is human IGF-I or an active fragment thereof.
20 . The method of claim 19 , comprising contacting said neuronal cells with human IGF-I.
21 . The method of claim 1 , wherein step (b) comprises contacting said neuronal cells with an IGF analog.
22 . The method of claim 21 , wherein said IGF analog is a peptidomimetic.
23 . The method of claim 21 , wherein said IGF analog is a small molecule.
24 . The method of claim 21 , wherein said IGF analog is an IGF-I analog.
25 . The method of claim 1 , wherein said IGF or active fragment or analog thereof has at least 10-fold higher affinity for the IGF-I receptor than native human IGF-I.
26 . The method of claim 1 , wherein said IGF or active fragment or analog thereof has an altered affinity for an IGF-binding protein (IBP).
27 . The method of claim 1 , wherein said IGF or active fragment or analog thereof has a half-life greater than native human IGF.
28 . The method of claim 1 , wherein step (a) and step (b) are in vitro.
29 . The method of claim 1 , wherein step (a) and step (b) are in vivo.
30 . A method of preventing or reducing the severity of an acute neurologic condition in a subject, comprising:
(a) administering to said subject EPO or an active fragment or analog thereof close to or subsequent to the time of acute injury; and
(b) administering to said subject an IGF or an active fragment or analog thereof close to or subsequent to the time of acute injury,
thereby providing a synergistic acute neuroprotective effect and preventing or reducing the severity of the acute neurologic condition.
31 . The method of claim 30 , wherein said acute neurologic condition is stroke.
32 . The method of claim 30 , wherein said acute neurologic condition is head or spinal cord trauma.
33 . The method of claim 30 , wherein said acute neurologic condition is seizure.
34 . The method of claim 30 , wherein step (a) comprises administering EPO or an active fragment thereof.
35 . The method of claim 34 , wherein said EPO is human EPO or an active fragment thereof.
36 . The method of claim 35 , comprising administering human EPO.
37 . The method of claim 30 , wherein step (a) comprises administering an EPO analog.
38 . The method of claim 37 , wherein said EPO analog is selected from the group consisting of GGTYSCHFGPLTWVCKPQGG (SEQ ID NO: 7);
GGDYHCRMGPLTWVCKPLGG (SEQ ID NO: 8);
GGVYACRMGPITWVCSPLGG (SEQ ID NO: 9);
VGNYMCHFGPITWVCRPGGG (SEQ ID NO: 10);
GGLYLCRFGPVTWDCGYKGG (SEQ ID NO: 11); and
GGCRIGPITWVCGG (SEQ ID NO: 12).
39 . The method of claim 37 , wherein said EPO analog is a peptidomimetic.
40 . The method of claim 37 , wherein said EPO analog is a small molecule.
41 . The method of claim 30 , wherein said EPO or active fragment or analog thereof has at least 10-fold higher affinity for the EPO receptor than native human EPO.
42 . The method of claim 30 , wherein said EPO or active fragment or analog thereof is oligomeric.
43 . The method of claim 42 , wherein said oligomeric EPO or active fragment or analog thereof is dimeric.
44 . The method of claim 43 , wherein said dimeric EPO or active fragment or analog thereof comprises dimeric GGTYSCHFGPLTWVCKPQGG (SEQ ID NO: 7).
45 . The method of claim 30 , wherein said EPO or active fragment or analog thereof has a half-life greater than native human EPO.
46 . The method of claim 30 , wherein said EPO or active fragment or analog thereof is hyper-glycosylated compared to native human EPO.
47 . The method of claim 30 , wherein step (a) comprises administering Darbepoietin.
48 . The method of claim 30 , further comprising administering soluble EPO receptor.
49 . The method of claim 30 , wherein step (b) comprises administering an IGF or an active fragment thereof.
50 . The method of claim 49 , wherein said IGF is IGF-I or an active fragment thereof.
51 . The method of claim 50 , wherein said IGF-I is human IGF-I or an active fragment thereof.
52 . The method of claim 50 , wherein said IGF-I is human IGF-I.
53 . The method of claim 30 , wherein step (b) comprises administering an IGF analog.
54 . The method of claim 53 , wherein said IGF analog is a peptidomimetic.
55 . The method of claim 53 , wherein said IGF analog is a small molecule.
56 . The method of claim 53 , wherein said IGF analog is an IGF-I analog.
57 . The method of claim 30 , wherein said IGF or active fragment or analog thereof has at least 10-fold higher affinity for the IGF-I receptor than native human IGF-I.
58 . The method of claim 30 , wherein said IGF or active fragment or analog thereof has an altered affinity for an IGF-binding protein (IBP).
59 . The method of claim 30 , wherein said IGF or active fragment or analog thereof has a half-life greater than native human IGF.
60 . A method of preventing or reducing the severity of a neurologic condition in a subject, comprising:
(a) administering to said subject EPO or an active fragment or analog thereof at a dose of at most 2000 U/kg; and
(b) administering to said subject an IGF or an active fragment or analog thereof,
thereby providing neuroprotection and preventing or reducing the severity of the neurologic condition.
61 . The method of claim 60 , wherein said neurologic condition is selected from the group consisting of 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 and anxiety.
62 . The method of claim 60 , wherein step (a) comprises administering EPO or an active fragment thereof.
63 . The method of claim 62 , wherein said EPO is human EPO or an active fragment thereof.
64 . The method of claim 63 , comprising administering human EPO.
65 . The method of claim 60 , wherein step (a) comprises administering an EPO analog.
66 . The method of claim 65 , wherein said EPO analog is selected from the group consisting of GGTYSCHFGPLTWVCKPQGG (SEQ ID NO: 7);
GGDYHCRMGPLTWVCKPLGG (SEQ ID NO: 8);
GGVYACRMGPITWVCSPLGG (SEQ ID NO: 9);
VGNYMCHFGPITWVCRPGGG (SEQ ID NO: 10);
GGLYLCRFGPVTWDCGYKGG (SEQ ID NO: 11); and
GGCRIGPITWVCGG (SEQ ID NO: 12).
67 . The method of claim 65 , wherein said EPO analog is a peptidomimetic.
68 . The method of claim 65 , wherein said EPO analog is a small molecule.
69 . The method of claim 60 , wherein said EPO or active fragment or analog thereof has at least 10-fold higher affinity for the EPO receptor than native human EPO.
70 . The method of claim 60 , wherein said EPO or active fragment or analog thereof is oligomeric.
71 . The method of claim 70 , wherein said oligomeric EPO or active fragment or analog thereof is dimeric.
72 . The method of claim 71 , wherein said dimeric EPO or active fragment or analog thereof comprises dimeric GGTYSCHFGPLTWVCKPQGG (SEQ ID NO: 7).
73 . The method of claim 60 , wherein said EPO or active fragment or analog thereof has a half-life greater than native human EPO.
74 . The method of claim 60 , wherein said EPO or active fragment or analog thereof is hyper-glycosylated compared to native human EPO.
75 . The method of claim 74 , wherein step (a) comprises administering Darbepoietin.
76 . The method of claim 60 , further comprising administering soluble EPO receptor.
77 . The method of claim 60 , wherein step (b) comprises administering an IGF or an active fragment thereof.
78 . The method of claim 77 , wherein said IGF is IGF-I or an active fragment thereof.
79 . The method of claim 78 , wherein said IGF-I is human IGF-I or an active fragment thereof.
80 . The method of claim 79 , wherein said IGF-I is human IGF-I.
81 . The method of claim 60 , wherein step (b) comprises administering an IGF analog.
82 . The method of claim 81 , wherein said IGF analog is a peptidomimetic.
83 . The method of claim 81 , wherein said IGF analog is a small molecule.
84 . The method of claim 81 , wherein said IGF analog is an IGF-I analog.
85 . The method of claim 60 , wherein said IGF or active fragment or analog thereof has at least 10-fold higher affinity for the IGF-I receptor than native human IGF-I.
86 . The method of claim 60 , wherein said IGF or active fragment or analog thereof has an altered affinity for an IGF-binding protein (IBP).
87 . The method of claim 60 , wherein said IGF or active fragment or analog thereof has a half-life greater than native human IGF.
88 - 113 . (canceled)