Methods and compositions for producing a neurosalutary effect in a subject

Abstract

Methods and compositions for producing a neurosalutary effect in a subject are provided. These methods generally involve administering to a subject a therapeutically effective amount of a compound that modulates the activity of N-kinase, or analog thereof. Pharmaceutical and packaged formulations including the compounds of the invention, e.g., compounds that modulate the activity of N-kinase, are also provided.

We claim:  
     
         1 . A method comprising administering to a subject a therapeutically effective amount of a compound that modulates the activity of N-kinase, thereby producing a neurosalutary effect in said subject.  
     
     
         2 . The method of  claim 1 , wherein the neurosalutary effect is produced in said subject by modulating neuronal survival.  
     
     
         3 . The method of  claim 1 , wherein the neurosalutary effect is produced in said subject by modulating neuronal regeneration.  
     
     
         4 . The method of  claim 1 , wherein the neurosalutary effect is produced in said subject by modulating neuronal axonal outgrowth.  
     
     
         5 . The method of  claim 1 , wherein the neurosalutary effect is produced in said subject by modulating axonal outgrowth of central nervous system neurons.  
     
     
         6 . The method of  claim 5 , wherein the central nervous system neurons are retinal ganglion cells.  
     
     
         7 . The method of  claim 1 , wherein the compound that modulates the activity of N-kinase is administered by introduction into a region of neuronal injury.  
     
     
         8 . The method of  claim 1 , wherein the compound that modulates the activity of N-kinase is introduced into the cerebrospinal fluid of the subject.  
     
     
         9 . The method of  claim 1 , wherein the compound that modulates the activity of N-kinase is introduced to the subject intrathecally.  
     
     
         10 . The method of  claim 1 , wherein the compound that modulates the activity of N-kinase is introduced into a region selected from the group consisting of a cerebral ventricle, the lumbar area, and the cisterna magna of the subject.  
     
     
         11 . The method of  claim 1 , wherein the compound that modulates the activity of N-kinase is administered to the subject in a pharmaceutically acceptable formulation.  
     
     
         12 . The method of  claim 11 , wherein the pharmaceutically acceptable formulation is a dispersion system.  
     
     
         13 . The method of  claim 11 , wherein the pharmaceutically acceptable formulation comprises a lipid-based formulation.  
     
     
         14 . The method of  claim 13 , wherein the pharmaceutically acceptable formulation comprises a liposome formulation.  
     
     
         15 . The method of  claim 13 , wherein the pharmaceutically acceptable formulation comprises a multivesicular liposome formulation.  
     
     
         16 . The method of  claim 11 , wherein the pharmaceutically acceptable formulation comprises a polymeric matrix.  
     
     
         17 . The method of  claim 11 , wherein the pharmaceutically acceptable formulation is contained within a minipump.  
     
     
         18 . The method of  claim 11 , wherein the pharmaceutically acceptable formulation provides sustained delivery of the compound that modulates the activity of N-kinase, to a subject for at least one week after the pharmaceutically acceptable formulation is administered to the subject.  
     
     
         19 . The method of  claim 13 , wherein the pharmaceutically acceptable formulation provides sustained delivery of the compound that modulates the activity of N-kinase, to a subject for at least one month after the pharmaceutically acceptable formulation is administered to the subject.  
     
     
         20 . The method of  claim 1 , wherein the subject is a mammal.  
     
     
         21 . The method of  claim 20 , wherein the mammal is a human.  
     
     
         22 . The method of  claim 1 , wherein said subject is suffering from a neurological disorder.  
     
     
         23 . The method of  claim 22 , wherein said neurological disorder is a spinal cord injury.  
     
     
         24 . The method of  claim 23 , wherein the spinal cord injury is characterized by monoplegia, diplegia, paraplegia, hemiplegia and quadriplegia.  
     
     
         25 . The method of  claim 22 , wherein said neurological disorder is epilepsy.  
     
     
         26 . The method of  claim 22 , wherein said neurological disorder is stroke.  
     
     
         27 . The method of  claim 22 , wherein said neurological disorder is Alzheimer's disease.  
     
     
         28 . A method comprising administering a therapeutically effective amount of a compound that modulates the activity of N-kinase to a subject suffering from a neurological disorder, thereby treating said subject suffering from a neurological disorder.  
     
     
         29 . The method of  claim 28 , further comprising making a first assessment of a nervous system function prior to administering the compound that modulates the activity of N-kinase to the subject and making a second assessment of the nervous system function after administering the compound that modulates the activity of N-kinase to the subject.  
     
     
         30 . The method of  claim 29 , wherein the nervous system function is a sensory function, cholinergic innervation, or a vestibulomotor function.  
     
     
         31 . A method for identifying a compound capable of producing a neurosalutary effect in a subject, comprising contacting N-kinase, or a biologically active fragment thereof, with a test compound and determining the ability of the test compound to modulate the activity of N-kinase, thereby identifying a compound capable of producing a neurosalutary effect in a subject.  
     
     
         32 . The method of  claim 31 , wherein the N-kinase is human N-kinase.  
     
     
         33 . The method of  claim 32 , wherein the human N-kinase is a recombinantly produced N-kinase.  
     
     
         34 . The method of  claim 31 , wherein the N-kinase is bovine N-kinase.  
     
     
         35 . The method of  claim 34 , wherein the bovine N-kinase is purified from a bovine source.  
     
     
         36 . The method of  claim 31 , further comprising determining the ability of the test compound to modulate axonal outgrowth of a central nervous system neuron.  
     
     
         37 . The method of  claim 31 , wherein the test compound inhibits the activity of N-kinase.  
     
     
         38 . The method of  claim 31 , wherein the test compound stimulates the activity of N-kinase.  
     
     
         39 . The method of  claim 31 , wherein the ability of the test compound to modulate the activity of N-kinase is determined by assessing the ability of the test compound to modulate N-kinase dependent phosphorylation of a substrate.  
     
     
         40 . A method for identifying a compound capable of producing a neurosalutary effect in a subject, comprising 
 contacting N-kinase or a biologically active fragment thereof, with a test compound, an N-kinase substrate, radioactive ATP, and Mn +2 ; and    determining the ability of the test compound to modulate N-kinase dependent phosphorylation of the substrate, thereby identifying a compound capable of producing a neurosalutary effect in a subject.    
     
     
         41 . The method of  claim 40 , wherein the N-kinase substrate is a histone HF-1 protein.  
     
     
         42 . The method of  claim 40 , wherein the radioactive ATP is [γ- 32 P] ATP.  
     
     
         43 . The method of  claim 40 , wherein the N-kinase is human N-kinase.  
     
     
         44 . The method of  claim 43 , wherein the human N-kinase is a recombinantly produced N-kinase.  
     
     
         45 . The method of  claim 40 , wherein the N-kinase is bovine N-kinase.  
     
     
         46 . The method of  claim 45 , wherein the bovine N-kinase is purified from a bovine source.  
     
     
         47 . The method of  claim 40 , further comprising determining the ability of the test compound to modulate axonal outgrowth of a central nervous system neuron.  
     
     
         48 . A compound capable of producing a neurosalutary effect in a subject identified by the method of  claim 40 .  
     
     
         49 . An isolated N-kinase polypeptide of the type that: 
 (a) is present in neonatal brain tissue;    (b) is inhibited in the presence of 6-thioguanine;    (c) is activated in the presence of Mn +2  but not by Mg +2  or Ca +2 ;    (d) has a molecular weight of approximately 49 kDa; and    (e) is eluted from a Cibacron Blue column at a NaCl concentration of 1.5-1.75 M.    
     
     
         50 . An antibody which is specifically reactive with an epitope of the N-kinase polypeptide of  claim 49 .  
     
     
         51 . The antibody of  claim 50 , wherein the antibody is an intracellular antibody.  
     
     
         52 . The antibody of  claim 50 , wherein the epitope comprises an ATP binding domain.  
     
     
         53 . A fragment of the N-kinase polypeptide of  claim 49 , wherein the fragment comprises at least 15 contiguous amino acids.  
     
     
         54 . The fragment of  claim 53 , wherein the fragment comprises at least 30 contiguous amino acids.  
     
     
         55 . The fragment of  claim 53 , wherein the fragment comprises at least 50 contiguous amino acids.  
     
     
         56 . The fragment of  claim 53 , wherein the fragment comprises at least 100 contiguous amino acids.  
     
     
         57 . A fragment of the N-kinase polypeptide of  claim 49 , wherein the fragment is able to elicit an immune response.  
     
     
         58 . An isolated nucleic acid molecule that encodes the polypeptide of SEQ ID NO:1.