Neuroprotective synergy of erythropoietin and insulin-like growth factors

Abstract

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)