Inhibitors of hedgehog signaling pathways, compositions and uses related thereto

Abstract

The present invention makes availables assays and reagents inhibiting paracrine and/or autocrine signals produced by a hedgehog protein or aberrant activation of a hedgehog signal transduction pathway, e.g., which involve the use of a steroidal alkaloid or other small molecule.

1 - 26 . (canceled)  
     
     
         27 . An in vitro method for inhibiting mitotic cell proliferation in a cell, comprising contacting the cell in vitro with a compound represented in the general formula (I), or unsaturated forms thereof, or pharmaceutically acceptable salts thereof:  
       
         
           
           
               
               
           
         
       
       wherein, as valence permits, 
 R 2 , R 3 , R 4 , and R 5  independently for each occurrence, represent one or more substituents selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═O, ═S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 ;  
 R 6 , R 7 , and R′ 7  are absent or represent, independently for each occurrence, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═O, ═S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 , or  
 R 6  and R 7 , or R 7  and R′ 7 , taken together form a ring or polycyclic ring;  
 with the proviso that at least one of R 6 , R 7 , or R′ 7  is present and includes a primary or secondary amine;  
 R 8  represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocycle, or a polycycle; and  
 m is an integer in the range 0 to 8 inclusive.  
 
     
     
         28 . An in vitro method for inhibiting mitotic cell proliferation in a cell, comprising contacting the cell in vitro with a compound represented in the general formula (II), or unsaturated forms thereof, or pharmaceutically acceptable salts thereof:  
       
         
           
           
               
               
           
         
         wherein, as valence permits,  
         R 2  and R 4 , independently for each occurrence, are selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═O, ═S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 ;  
         R 3 , independently for each occurrence, is selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 ;  
         R 5 , independently for each occurrence, is absent or represents a substituent selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 ;  
         R 6 , R 7 , and R′ 7  are absent or represent, independently, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═O, ═S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 , or  
         R 6  and R 7 , or R′ 7  and R′ 7 , taken together form a ring or polycyclic ring,  
         with the proviso that at least one of R 6 , R 7 , or R′ 7  is present and includes a primary or secondary amine;  
         R 8  represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocycle, or a polycycle;  
         X represents O or S; and  
         m is an integer in the range 0 to 8 inclusive.  
       
     
     
         29 . The method of any one of claims  27 - 28 , wherein the cell is a hyperproliferative cell.  
     
     
         30 . The method of  claim 29 , wherein the hyperproliferative cell is a cancer cell.  
     
     
         31 . The method of  claim 30 , wherein the cancer is a basal cell carcinoma, medulloblastoma, squamous cell carcinoma, carcinosarcoma, adenocystic carcinoma, epidermoid carcinoma, nasopharyngeal carcinoma, renal cell carcinoma, papilloma, or an epidermoidoma.  
     
     
         32 . The method of any one of claims  27 - 28 , wherein the compound is jervine or cyclopamine.  
     
     
         33 . The method of any one of claims  27 - 28 , wherein the compound inhibits hedgehog-mediated signal transduction with an ED 50  of 1 mM or less.  
     
     
         34 . The method of any one of claims  27 - 28 , wherein the compound inhibits hedgehog-mediated signal transduction with an ED 50  of 1 μM or less.  
     
     
         35 . The method of any one of claims  27 - 28 , wherein the compound inhibits hedgehog-mediated signal transduction with an ED 50  of 1 nM or less.  
     
     
         36 . The method of any one of claims  27 -28, wherein R 2  and R 3 , independently for each occurrence, are —OH, alkyl, —O-alkyl, —C(O)-alkyl, or —C(O)—R 8 .  
     
     
         37 . The method of any one of claims  27 - 28 , wherein R 4 , independently for each occurrence represents —OH, ═O, alkyl, —O-alkyl, —C(O)-alkyl, or —C(O)—R 8 .  
     
     
         38 . The method of any one of claims  27 - 28 , wherein R 6 , R 7 , and R′ 7 , independently for each occurrence, represent hydrogen, alkyls, alkenyls, alkynyls, amines, imines, amides, carbonyls, carboxyls, carboxamides, ethers, thioethers, esters, or —(CH 2 ) m —R 8 .  
     
     
         39 . The method of any one of claims  27 - 28 , wherein R 7 , and R′ 7  taken together form a substituted or unsubstituted furanopiperidine.  
     
     
         40 . The method of  claim 39 , wherein the furanopiperidine is a substituted or unsubstituted perhydrofuro[3,2-b]pyridine, a pyranopiperidine, a quinoline, an indole, a pyranopyrrole, a naphthyridine, a thiofuranopiperidine, or a thiopyranopiperidine.  
     
     
         41 . The method of  claim 27 , wherein the compound is represented in Formula (Ia), or unsaturated forms thereof, or pharmaceutically acceptable salts thereof:  
       
         
           
           
               
               
           
         
       
       wherein, as valence permits, 
 R 2 , R 3 , R 4 , and R 5  independently for each occurrence, represent one or more substituents selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═O, ═S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 ;  
 R 6 , R 7 , and R′ 7  are absent or represent, independently for each occurrence, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═O, ═S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 , or  
 R6 and R 7 , or R′ 7  and R′ 7 , taken together form a ring or polycyclic ring; with the proviso that at least one of R6, R 7 , or R′ 7  is present and includes a primary or secondary amine;  
 R 8  represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocycle, or a polycycle; and m is an integer in the range 0 to 8 inclusive.  
 
     
     
         42 . The method of  claim 28 , wherein the compound is represented in Formula (IIa), or unsaturated forms thereof, or pharmaceutically acceptable salts thereof:  
       
         
           
           
               
               
           
         
       
       wherein, as valence permits, 
 R 2  and R 4 , independently for each occurrence, are selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═O, ═S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes., esters, or —(CH 2 ) m —R 8 ;  
 R 3 , independently for each occurrence, are selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 ;  
 R 5 , independently for each occurrence, is absent or is selected from hydrogen, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 ;  
 R 6 , R 7 , and R′ 7  are absent or represent, independently, halogens, alkyls, alkenyls, alkynyls, aryls, hydroxyl, ═O, ═S, alkoxyl, silyloxy, amino, nitro, thiol, amines, imines, amides, phosphoryls, phosphonates, phosphines, carbonyls, carboxyls, carboxamides, anhydrides, silyls, ethers, thioethers, alkylsulfonyls, arylsulfonyls, selenoethers, ketones, aldehydes, esters, or —(CH 2 ) m —R 8 , or  
 R 6  and R 7 , or R 7  and R′ 7 , taken together form a ring or polycyclic ring, with the proviso that at least one of R 6 , R 7 , or R′ 7  is present and includes a primary or secondary amine;  
 R 8  represents an aryl, a cycloalkyl, a cycloalkenyl, a heterocycle, or a polycycle;  
 X represents O or S; and  
 m is an integer in the range 0 to 8 inclusive.