Mediators of hedgehog signaling pathways, compositions and uses related thereto

Abstract

The present invention makes available methods and reagents for inducing activation of a hedgehog pathway in a cell or tissue comprising contacting the cell with a hedgehog agonist, such as a small molecule, in a sufficient amount to achieve a therapeutic result. In certain embodiments, the methods and reagents may be employed to correct or inhibit an aberrant or unwanted growth state, e.g., by antagonizing a normal ptc pathway or agonizing smoothened or hedgehog activity.

1 . A compound represented in general formula (III):  
       
         
           
           
               
               
           
         
       
       wherein 
 Ar and Ar′ independently represent substituted or unsubstituted aryl or heteroaryl rings;  
 Y, independently for each occurrence, is absent or represents —NR—, —O—, —S—, or —Se—;  
 X is selected from —C(═O)—, —C(═S)—, —S(O 2 )—, —S(O)—, —C(═NCN)—, —P(═O)(OR 2 )—, and a methylene group optionally substituted with 1-2 groups selected from lower alkyl, alkenyl, and alkynyl groups;  
 M represents, independently for each occurrence, a substituted or unsubstituted methylene group, or two adjacent M taken together represent substituted or unsubstituted ethene or ethyne:  
 R represents, independently for each occurrence, H or substituted or unsubstituted aryl, heterocyclyl, heteroaryl, aralkyl, heteroaralkyl, alkynyl, alkenyl, or alkyl, or two R taken together may form a 4- to 8-membered ring,  
 Cy represents a cyclohexyl ring,  
 Cy′ represents a substituted or unsubstituted aryl, heterocyclyl, heteroaryl, or cycloalkyl; and  
 i represents, independently for each occurrence, an integer from 0 to 5;  
 wherein Y-M i  adjacent to Cy is absent for both occurrences, and the compound is enriched for the isomer in which the two nitrogen substituents on Cy are disposed in a trans-relationship.  
 
     
     
         2 . The compound of  claim 1 , wherein Y is absent from all positions.  
     
     
         3 . The compound of  claim 1 , wherein M represents, independently for each occurrence, a substituted or unsubstituted methylene group.  
     
     
         4 . The compound of  claim 1 , wherein X is selected from —C(═O)—, —C(═S)—, and —S(O 2)—.    
     
     
         5 . The compound of  claim 1 , wherein NR 2  represents a primary amine, or a secondary amine substituted with one lower alkyl group.  
     
     
         6 . The compound of  claim 1 , wherein Cy′ is a substituted or unsubstituted aryl or heteroaryl.  
     
     
         7 . The compound of  claim 1 , wherein Cy′ is bicyclic.  
     
     
         8 . The compound of  claim 7 , wherein Cy′ represents a substituted or unsubstituted benzothiophene.  
     
     
         9 . The compound of  claim 8 , wherein the benzothiophene ring is substituted with one or more methyl, ethyl, fluorine, chlorine, methoxy or cyano groups.  
     
     
         10 . The compound of  claim 9 , wherein each R independently represents H or a lower alkyl.  
     
     
         11 . The compound of  claim 9 , wherein NR 2  represents a secondary amine.  
     
     
         12 . The compound of  claim 11 , wherein each R independently represents H or a lower alkyl.  
     
     
         13 . The compound of  claim 10 , wherein 
 i is zero except for M i  between Ar and N; and all Y are absent.    
     
     
         14 . The compound of  claim 12 , wherein 
 i is zero except for M i  between Ar and N; and all Y are absent.    
     
     
         15 . The compound of  claim 14 , wherein X is selected from —C(═O)—, —C(═S)—, and —S(O 2 )—.  
     
     
         16 . The compound of  claim 11 , wherein X is selected from —C(═O)—, —C(═S)—, and —S(O 2 )—.  
     
     
         17 . The compound of  claim 10 , wherein X is selected from —C(═O)—, —C(═S)—, and —S(O 2 )—.  
     
     
         18 . The compound of  claim 8 , wherein X is selected from —C(═O)—, —C(═S)—, and —S(O 2 )—.  
     
     
         19 . The compound of  claim 18 , wherein NR 2  represents a secondary amine.  
     
     
         20 . The compound of  claim 19 , wherein each R independently represents H or a lower alkyl.  
     
     
         21 . The compound of  claim 8 , wherein NR 2  represents a secondary amine.  
     
     
         22 . The compound of  claim 20 , wherein 
 i is zero except for M i  between Ar and N; and    all Y are absent.    
     
     
         23 . The compound of  claim 21 , wherein 
 i is zero except for M, between Ar and N, and    all Y are absent.    
     
     
         24 . The compound of  claim 23 , wherein each R independently represents H or a lower alkyl.  
     
     
         25 . The compound of  claim 24 , wherein NR 2  represents a secondary amine.  
     
     
         26 . The compound of  claim 25 , wherein Cy′ represents a substituted or unsubstituted benzothiophene.  
     
     
         27 . The compound of  claim 26 , wherein X is selected from —C(═O)—, —C(═S)—, and —S(O 2 )—.  
     
     
         28 . The compound of  claim 27 , wherein the benzothiophene ring is substituted with one or more methyl, ethyl, fluorine, chlorine, methoxy or cyano groups.  
     
     
         29 . The compound of  claim 27 , wherein the benzothiophene ring is substituted with one or more methyl, fluorine, or chlorine groups.  
     
     
         30 . A compound represented in general formula (Vet):  
       
         
           
           
               
               
           
         
       
       wherein 
 Cy represents a cyclohexyl ring;  
 Ar is a substituted or unsubstituted aryl or heteroaryl ring;  
 W is O or S,  
 R represents, independently for each occurrence, H or substituted or unsubstituted aryl, heterocyclyl, heteroaryl, aralkyl, heteroaralkyl, alkynyl, alkenyl, or alkyl, or two R taken together may form a 4- to 8-membered ring;  
 R 1  and R 2  represent, independently and as valency permits, from 0-5 substituents on the ring to which each is attached, selected from halogen, lower alkyl, lower alkenyl, aryl, heteroaryl, carbonyl, thiocarbonyl, ketone, aldehyde, amino, acylamino, amido, amidino, cyano, nitro, hydroxyl, azido, sulfonyl, sulfoxido, sulfate, sulfonate, sulfamoyl, sulfonamido, phosphoryl, phosphonate, phosphinate, —(CH 2 ) p alkyl, —(CH 2 ) p alkenyl, —(CH 2 ) p alkynyl, —(CH 2 ) p aryl, —(CH 2 ) p aralkyl, —(CH 2 ) p OH, —(CH 2 ) p O-lower alkyl, —(CH 2 ) p O-lower alkenyl, —O(CH 2 ) n R, —(CH 2 ) p SH, —(CH 2 ) p S-lower alkyl, —(CH 2 ) p S-lower alkenyl, —S(CH 2 ) n R, —(CH 2 ) p NR 2 , —(CH 2 ) p NR-lower alkyl, —(CH 2 ) p NR-lower alkenyl, —NR(CH 2 ) n R, and protected forms of the above; and  
 n and p, individually for each occurrence, represent integers from 0 to 10,  
 wherein the compound is enriched for the isomer in which the two nitrogen substituents on Cy are disposed in a trans-relationship.  
 
     
     
         31 . The compound of  claim 30 , wherein Ar is heteroaryl.  
     
     
         32 . The compound of  claim 30 , wherein Ar is bicyclic.  
     
     
         33 . The compound of  claim 30 , wherein NR 2  represents NH).  
     
     
         34 . The compound of  claim 33 , wherein Ar represents a substituted or unsubstituted benzothiophene.  
     
     
         35 . The compound of  claim 34 , wherein the benzothiophene ring is substituted with one or more methyl, ethyl, fluorine, chlorine, methoxy, or cyano groups.  
     
     
         36 . The compound of  claim 30 , wherein each R independently represents H or a lower alkyl group.  
     
     
         37 . The compound of  claim 36 , wherein NR 2  represents a secondary amine.  
     
     
         38 . The compound of  claim 36 , wherein Ar represents a substituted or unsubstituted benzothiophene.  
     
     
         39 . The compound of  claim 38 , wherein the benzothiophene ring is substituted with one or more methyl, ethyl, fluorine, chlorine, methoxy, or cyano groups.  
     
     
         40 . The compound of  claim 30 , wherein Ar represents a substituted or unsubstituted benzothiophene.  
     
     
         41 . The compound of  claim 40 , wherein the benzothiophene ring is substituted with one or more methyl, ethyl, fluorine, chlorine, methoxy, or cyano groups.  
     
     
         42 . The compound of  claim 40 , wherein each R independently represents H or a lower alkyl group.  
     
     
         43 . The compound of  claim 42 , wherein NR 2  represents a secondary amine.  
     
     
         44 . The compound of  claim 43 , wherein the benzothiophene ring is substituted with one or more methyl, ethyl, fluorine, chlorine, methoxy, or cyano groups.  
     
     
         45 . The compound of  claim 43 , wherein the benzothiophene ring is substituted with one or more methyl, fluorine, or chlorine groups.  
     
     
         46 . A method for modulating proliferation or differentiation of a cell, comprising contacting the cell with a compound of  claim 1  or  claim 30 .  
     
     
         47 . The method of  claim 46 , wherein the compound is a hedgehog agonist.  
     
     
         48 . The method of  claim 47 , wherein the hedgehog agonist agonizes hedgehog mediated signal transduction with an ED 50  of 1 μM or less.  
     
     
         49 . The method of  claim 47 , wherein the hedgehog agonist is administered as part of a therapeutic or cosmetic application.  
     
     
         50 . The method of  claim 49 , wherein the therapeutic or cosmetic application is selected from regulation of neural tissues, bone and cartilage formation and repair, regulation of spermatogenesis, regulation of smooth muscle, regulation of lung, liver and other organs arising from the primitive gut, regulation of hematopoietic function, and regulation of skin and hair growth.  
     
     
         51 . A method for treating neurological conditions deriving from acute, subacute, or chronic injury to the nervous system: aging of the nervous system; chronic neurodegenerative diseases of the nervous system; or chronic immunological diseases of the nervous system or affecting the nervous system, comprising administering to a subject the preparation of  claim 1  or  30 .  
     
     
         52 . The method of  claim 51 , wherein the injury is ischemic injury resulting from stroke.  
     
     
         53 . The method of  claim 51 , wherein the neurodegenerative disease is Parkinson's disease.  
     
     
         54 . The method of  claim 51 , wherein the neuromuscular disorders are selected from chronic or progressive atrophies and chronic peripheral neuropathy.  
     
     
         55 . A method for treating or reducing the risk of developing Parkinson's disease comprising administering to an animal a compound of  claim 1  or  30 .  
     
     
         56 . The method of  claim 55 , wherein the hedgehog agonist increases the rate of survival of dopaminergic neurons.  
     
     
         57 . A method for treating autonomic disorders of the peripheral nervous system comprising administering to an animal a compound of  claim 1  or  30 .  
     
     
         58 . A method for treating neuronal damage due to ischemic events comprising administering to an animal a compound of  claim 1  or  30 .  
     
     
         59 . A method of modulating the growth, differentiation, or survival of a cell, comprising contacting the cell with a compound of  claim 1  or  30 .  
     
     
         60 . The method of  claim 59 , wherein the cell is contacted with the compound in vitro.  
     
     
         61 . The method of  claim 59 , wherein the cell is contacted with the compound ex vivo.