Neuroprotective methods and reagents

Abstract

One aspect of the present application relates to a method for limiting damage to neuronal cells by ischemic or epoxic conditions, e.g., such as may be manifest by a reduction in brain infarct volume, by administering to an individual a hedgehog therapeutic or ptc therapeutic in an amount effective for reducing cerebral infarct volume.

1 . A method for limiting damage to neuronal cells by ischemic or epoxic conditions, comprising administering to an individual a ptc therapeutic in an amount effective for reducing cerebral infarct volume relative to the absence of administration of the ptc therapeutic, wherein the ptc therapeutic inhibits PKC with a K i  greater than 1 μM.  
     
     
         2 . A method for protecting cerebral tissue of a mammal against the repercussions of ischemia which comprises administering to the mammal in need thereof a therapeutically effective amount of a ptc therapeutic therapeutic, wherein the ptc therapeutic inhibits PKC with a K i  greater than 1 μM.  
     
     
         3 . A method for the treatment of cerebral infarctions which comprises administering to a patient in need thereof a therapeutically effective amount of a ptc therapeutic therapeutic, wherein the ptc therapeutic inhibits PKC with a K i  greater than 1 μM.  
     
     
         4 . A method for the treatment of cerebral ischemia which comprises administering to a patient in need thereof a therapeutically effective amount of a ptc therapeutic therapeutic, wherein the ptc therapeutic inhibits PKC with a K i  greater than 1 μM.  
     
     
         5 . A method for the treatment of stroke which comprises administering to a patient in need thereof a therapeutically effective amount of a ptc therapeutic therapeutic, wherein the ptc therapeutic inhibits PKC with a K i  greater than 1 μM.  
     
     
         6 . A method for the treatment of transient ischemia attack which comprises administering to a patient in need thereof a therapeutically effective amount of a ptc therapeutic therapeutic, wherein the ptc therapeutic inhibits PKC with a K i  greater than 1 μM.  
     
     
         7 . The method of  claim 1 , wherein the ptc therapeutic binds to patched and mimics hedgehog-mediated patched signal transduction.  
     
     
         8 . The method of  claim 7 , wherein the ptc therapeutic is a small organic molecule.  
     
     
         9 . The method of  claim 7 , wherein the binding of the ptc therapeutic to patched results in upregulation of patched and/or gli expression.  
     
     
         10 . The method of  claim 8 , wherein the ptc therapeutic is a small organic molecule which interacts with neuronal cells to mimic hedgehog-mediated patched signal transduction.  
     
     
         11 . The method of  claim 1 , wherein the ptc therapeutic mimics hedgehog-mediated patched signal transduction by altering the localization, protein-protein binding and/or enzymatic activity of an intracellular protein involved in a patched signal pathway.  
     
     
         12 . The method of  claim 1 , wherein the ptc therapeutic alters the level of expression of a hedgehog protein, a patched protein or a protein involved in the intracellular signal transduction pathway of patched.  
     
     
         13 . The method of  claim 11 , wherein the ptc therapeutic is a small organic molecule which binds to patched and regulates patched-dependent gene expression.  
     
     
         14 . The method of  claim 11 , wherein the ptc therapeutic is an inhibitor of protein kinase A (PKA).  
     
     
         15 . The method of  claim 14 , wherein the PKA inhibitor is a 5-isoquinolinesulfonamide.  
     
     
         16 . The method of  claim 15 , wherein the PKA inhibitor is represented in the general formula:  
       
         
           
           
               
               
           
         
       
       wherein, 
 R 1  and R 2  each can independently represent hydrogen, and as valence and stability permit a lower alkyl, a lower alkenyl, a lower alkynyl, a carbonyl, a thiocarbonyl, an amino, an acylamino, an amido, a cyano, a nitro, an azido, a sulfate, a sulfonate, a sulfonamido, —(CH 2 ) m —R 8 , —(CH 2 ) m —OH, —(CH 2 ) m —O-lower alkyl, —(CH 2 ) m —O-lower alkenyl, —(CH 2 ) n —O—(CH 2 ) m —R 8 , —(CH 2 ) m —SH, —(CH 2 ) m —S-lower alkyl, —(CH 2 ) m —S-lower alkenyl, —(CH 2 ) n —S—(CH 2 ) m —R 8 , or  
 R 1  and R 2  taken together with N form a substituted or unsubstituted heterocycle;  
 R 3  is absent or represents one or more substitutions to the isoquinoline ring such as a lower alkyl, a lower alkenyl, a lower alkynyl, a carbonyl (such as a carboxyl, an ester, a formate, or a ketone), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an amino, an acylamino, an amido, a cyano, a nitro, an azido, a sulfate, a sulfonate, a sulfonamido, —(CH 2 ) m —R 8 , —(CH 2 ) m —OH, —(CH 2 ) m —O-lower alkyl, —(CH 2 ) m —O-lower alkenyl, —(CH 2 ) n —O—(CH 2 ) m —R 8 , —(CH 2 ) m —SH, —(CH 2 ) m —S-lower alkyl, —(CH 2 ) m —S-lower alkenyl, —(CH 2 ) n —S—(CH 2 ) m —R 8 ;  
 R 8  represents a substituted or unsubstituted aryl, aralkyl, cycloalkyl, cycloalkenyl, or heterocycle; and  
 n and m are independently for each occurrence zero or an integer in the range of 1 to 6.  
 
     
     
         17 . The method of  claim 14 , wherein the PKA inhibitor is selected from the group consisting of N-[2-((p-bromocinnamyl)amino)ethyl]-5-isoquinolinesulfonamide, KT5720, and PKA Heat Stable Inhibitor isoform α.  
     
     
         18 . The method of  claim 5 , wherein the stroke is a thrombotic stroke.  
     
     
         19 . The method of  claim 5 , wherein the stroke is an embolic stroke.  
     
     
         20 . The method of  claim 1 , wherein the conditions result in cerebral hypoxia.  
     
     
         21 . The method of  claim 1 , wherein the conditions result in progressive loss of neurons due to oxygen deprivation.  
     
     
         22 . The method of  claim 3 , wherein the patient is treated prophylactically.  
     
     
         23 . The method of  claim 1 , wherein the individual is treated prophylactically.  
     
     
         24 . The method of  claim 2 , wherein the mammal is treated prophylactically.  
     
     
         25 . The method of  claim 1 , wherein the patient is hypotensive.  
     
     
         26 . The method of  claim 1 , further comprising administering one or more of an anticoagulant, an antiplatelet agent, a thrombin inhibitor, and/or a thrombolytic agent.  
     
     
         27 . The method of  claim 1 , further comprising performing vascular surgery.  
     
     
         28 . The method of  claim 27 , wherein the vascular surgery comprises carotid endarterectomy.  
     
     
         29 . The method of  claim 1 , wherein treatment of the patient with the ptc therapeutic results in at least a 25% reduction in cerebral infarct volumes relative to absence of treatment with the ptc therapeutic.  
     
     
         30 . The method of  claim 29 , wherein treatment of the patient with the ptc therapeutic results in at least a 50% reduction in cerebral infarct volumes relative to absence of treatment with the ptc therapeutic.  
     
     
         31 . The method of  claim 29 , wherein treatment of the patient with the ptc therapeutic results in at least a 70% reduction in cerebral infarct volumes relative to absence of treatment with the ptc therapeutic.  
     
     
         32 . The method of  claim 1 , wherein the ptc therapeutic inhibits the activity of PKA, cAMP, or adenylate cyclase.  
     
     
         33 . The method of  claim 1 , wherein the ptc therapeutc agonizes the activity of cAMP phosphodiesterase.  
     
     
         34 . A therapeutic preparation of a small molecule antagonist of patched, which patched antagonist inhibits PKC with a K 1  greater than 100 nM and is provided in a pharmaceutically acceptable carrier and in an amount sufficient to provide protection against neuronal cell death under ischemic and/or hypoxic conditions.  
     
     
         35 . The preparation of  claim 34 , which patched antagonist binds to patched.  
     
     
         36 . The preparation of  claim 34 , wherein the patched antagonist is provided in an amount sufficient to produce, upon a dosage regimen of 7 days, at least a 70% decrease in infarct volume in an MCAO model relative to the absence of the patched antagonist.  
     
     
         37 . The preparation of  claim 34 , wherein the patched antagonist is provided in an amount sufficient to produce, upon a dosage regimen of 3 days, at least a 70% decrease in infarct volume in an MCAO model relative to the absence of the patched antagonist.