Novel molecules for regulating cell death

Abstract

The present invention provides compounds capable of regulating apoptosis, e.g., via regulating mitochondrial fission or fusion. The present invention also provides methods of screening for compounds capable of regulating apoptosis and methods of treating conditions association with apoptosis.

1 . A compound of the general formula:  
       
         
           
           
               
               
           
         
         wherein R 1  is independently H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof;  
         R 2  is H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; or a halogen;  
         R 3  is H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof;  
         R 4  is H or a halogen; and  
         R 5  is H or a halogen,  
         with the provisos that when R 3  is H and R 4  is H or a halogen, R 5  is a halogen, or when R 2  is a halogen R 5  is H or a halogen,  
         further wherein said compound causes the formation of net like structures in mitochondria.  
       
     
     
         2 . The compound of  claim 1 , wherein R 2  is a halogen.  
     
     
         3 . The compound of  claim 2 , wherein R 2  is selected from F, Cl or Br.  
     
     
         4 . The compound of  claim 3 , wherein the compound is 3-(2-fluorophenyl)-2-mercaptoquinolin-4(3H)-one.  
     
     
         5 . The compound of  claim 3 , wherein the compound is 3-(2-chlorophenyl)-2-mercaptoquinolin-4(3H)-one.  
     
     
         6 . The compound of  claim 3 , wherein the compound is 3-(2-bromophenyl)-2-mercaptoquinolin-4(3H)-one.  
     
     
         7 . The compound of  claim 2 , wherein R 5  is a halogen.  
     
     
         8 . The compound of  claim 7 , wherein the compound is 3-(2,6-dichlorophenyl)-2-mercaptoquinolin-4(3H)-one.  
     
     
         9 . The compound of  claim 1 , wherein R 4  is H.  
     
     
         10 . The compound of  claim 9 , wherein the compound is 3-(6-chlorophenyl)-2-mercaptoquinazolin-4(3H)-one.  
     
     
         11 . The compound of  claim 1 , wherein R 4  is a halogen.  
     
     
         12 . The compound of  claim 11 , wherein the compound is 3-(2,4-dichlorophenyl)-2-mercaptoquinolin-4(3H)-one.  
     
     
         13 . The compound of  claim 12 , wherein R 3  is an alkyl which may be branched, may contain a heteroatom, may be substituted, or may contain a combination thereof.  
     
     
         14 . The compound of  claim 13 , wherein the compound is 3-(2,4-dichloro-5-methoxyphenyl)-2-mercaptoquinazolin-4(3H)-one.  
     
     
         15 . The compound of  claim 1 , wherein R 2  is an alkyl which may be branched, may contain a heteroatom, may be substituted, or may contain a combination thereof.  
     
     
         16 . The compound of  claim 15 , wherein the compound is 2-mercapto-3-O-tolyl-4(3H)-quinazolinone.  
     
     
         17 . The compound of  claim 1 , wherein R 2  is an alkyl which may be branched, may contain a heteroatom, may be substituted, or may contain a combination thereof.  
     
     
         18 . The compound of  claim 17 , wherein the compound is 2-mercapto-3-(2-(trifluoromethyl)phenyl)quinazolin-4(3H)-one.  
     
     
         19 . The compound of  claim 17 , wherein the compound is 2-mercapto-3-(2-ethyl-phenyl) quinazolin-4(3H)-one.  
     
     
         20 . A pharmaceutical composition comprising the compound of any one of claims  1 - 19  and a pharmaceutically acceptable carrier.  
     
     
         21 . A method of modulating mitochondrial fission in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  1 - 19 .  
     
     
         22 . A method of inhibiting apoptosis in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  1 - 19 .  
     
     
         23 . A method of treating a condition associated with apoptosis, wherein the treatment includes decreasing apoptosis comprising administering to a subject in need thereof a compound of any one of claims  1 - 19 .  
     
     
         24 . The method of  claim 23 , wherein the condition is selected from the group consisting of Parkinson's disease, ALS, stroke, heart attack, congestive heart failure, transplantation, alcoholic hepatitis, and drug-induced liver toxicity.  
     
     
         25 . A compound of the general formula:  
       
         
           
           
               
               
           
         
       
       wherein M is C and n is an integer of 0 or 1; 
 wherein R 1  is independently H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; 
 R 2  is H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; or a halogen;  
 R 3  is H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof;  
 R 4  is H or a halogen; and  
 R 5  is H or a halogen,  
 with the provisos that when R 3  is H and R 4  is H or a halogen, R 5  is a halogen, or when R 2  is a halogen R 5  is H or a halogen,  
 further wherein said compound causes the formation of net like structures in mitochondria.  
 
 
     
     
         26 . A pharmaceutical composition comprising the compound of  claim 20  and a pharmaceutically acceptable carrier.  
     
     
         27 . A method of modulating mitochondrial fission in a cell comprising contacting the cell with a formulation comprising the compound of  claim 20 .  
     
     
         28 . A method of inhibiting apoptosis in a cell comprising contacting the cell with a formulation comprising the compound of  claim 20 .  
     
     
         29 . A method of treating a condition associated with apoptosis, wherein the treatment includes decreasing apoptosis comprising administering to a subject in need thereof a compound of  claim 20 .  
     
     
         30 . The method of  claim 29 , wherein the condition is selected from the group consisting of Parkinson's disease, ALS, stroke, heart attack, congestive heart failure, transplantation, alcoholic hepatitis, and drug-induced liver toxicity.  
     
     
         31 . A compound of the general formula:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2  and R 3  are independently H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a halogen or a combination thereof,  
       further wherein said compound inhibits mitochondrial fusion.  
     
     
         32 . The compound of  claim 31 , wherein R 1 -R 3  are different.  
     
     
         33 . The compound of  claim 31 , wherein R 1 -R 3  are identical.  
     
     
         34 . The compound of  claim 33 , wherein R 1 -R 3  are halogens.  
     
     
         35 . The compound of  claim 34 , wherein the compound is 1,3-dinitro-5-(trifluoromethyl)-2-(4-(trifluoromethyl)phenyoxy)benzene.  
     
     
         36 . The compound of  claim 33 , wherein the compound is 5-tert-butyl-1,3-dinitro-2-(4-(trifluoromethyl)phenoxy)benzene.  
     
     
         37 . A pharmaceutical composition comprising the compound of any one of claims  31 - 36  and a pharmaceutically acceptable carrier.  
     
     
         38 . A method of modulating mitochondrial fusion in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  31 - 36 .  
     
     
         39 . A method of increasing apoptosis in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  31 - 36 .  
     
     
         40 . A method of treating a condition associated with apoptosis, wherein the treatment includes increasing apoptosis comprising administering to a subject in need thereof a compound of any one of claims  31 - 36 .  
     
     
         41 . The method of  claim 40 , wherein the condition is neoplasia.  
     
     
         42 . A compound of the general formula:  
       
         
           
           
               
               
           
         
       
       wherein W is H or O; 
 R 1  and R 2  are independently H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or may combine to form a C5-18 cycloalkyl;  
 R 3  is a H or a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof;  
 R 4  is a H or a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof;  
 R 5  is H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or may combine to form a C5-18 cycloalkyl,  
 further wherein said compound inhibits mitochondrial fusion.  
 
     
     
         43 . The compound of  claim 42 , wherein R 1  and R 2  form a cycloalkyl.  
     
     
         44 . The compound of  claim 43 , wherein cycloalkyl, is a cyclopentyl ring.  
     
     
         45 . The compound of  claim 44 , wherein the compound is 5-(6-tert-butyl-1,1-dimethyl-2,3-dihydro-1H-inden-4-yl)-3-(trifluoromethyl)-4,5-dihydro-1H-pyrazole.  
     
     
         46 . The compound of  claim 42 , wherein R 3  is a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof.  
     
     
         47 . The compound of  claim 46 , wherein R 4  is a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof.  
     
     
         48 . The compound of  claim 47 , wherein the compound is 5-(6-bromobenzoyl)-5-(4-ethylphenyl)-3-(trifluoromethyl)-4,5-dihydro-1H-pyrazol-5-olate.  
     
     
         49 . A pharmaceutical composition comprising the compound of any one of claims  42 - 48  and a pharmaceutically acceptable carrier.  
     
     
         50 . A method of modulating mitochondrial fusion in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  42 - 48 .  
     
     
         51 . A method of increasing apoptosis in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  42 - 48 .  
     
     
         52 . A method of treating a condition associated with apoptosis, wherein the treatment includes increasing apoptosis comprising administering to a subject in need thereof a compound of any one of claims  42 - 48 .  
     
     
         53 . The method of  claim 52 , wherein the condition is neoplasia.  
     
     
         54 . A compound of the general formula:  
       
         
           
           
               
               
           
         
       
       R 1  and R 2  are independently H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or a halogen; or combinations thereof; 
 R 3  is are independently H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or a combination thereof,  
 further wherein said compound inhibits mitochondrial fusion.  
 
     
     
         55 . The compound of  claim 54 , wherein R 3  is a C3-C18 aryl which may contain a side group.  
     
     
         56 . The compound of  claim 55 , wherein R 1  is a halogen.  
     
     
         57 . The compound of  claim 56 , wherein the halogen is Cl.  
     
     
         58 . The compound of  claim 57 , wherein the compound is N 5 -(4-chlorophenyl)-N-6-phenethyl-[1,2,5]oxadiazolo[3,4-b]pyrazine-5,6-diamine.  
     
     
         59 . The compound of  claim 54 , wherein R 1  and R 2  are identical.  
     
     
         60 . The compound of  claim 59 , wherein R 1  and R 2  are H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted.  
     
     
         61 . The compound of  claim 60 , wherein R 3  is a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted.  
     
     
         62 . The compound of  claim 61 , wherein the compound is N 5 -cyclohexyl-N-6-(3,4-dimethylohenyl)-[1,2,5]oxadiazolo[3,4-b]pyrazine-5,6-diamine.  
     
     
         63 . A pharmaceutical composition comprising the compound of any one of claims  54 - 62  and a pharmaceutically acceptable carrier.  
     
     
         64 . A method of modulating mitochondrial fusion in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  54 - 62 .  
     
     
         65 . A method of increasing apoptosis in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  54 - 62 .  
     
     
         66 . A method of treating a condition associated with apoptosis, wherein the treatment includes increasing apoptosis comprising administering to a subject in need thereof a compound of any one of claims  54 - 62 .  
     
     
         67 . The method of  claim 66 , wherein the condition is neoplasia.  
     
     
         68 . A compound of the general formula:  
       
         
           
           
               
               
           
         
       
       wherein 
 W is N or S;  
 R 1  is H or a C1-C3 alkyl or a substituted piperidine;  
 R 2  is H or a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or a combination thereof;  
 R 3  is H; O or a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or a combination thereof;  
 R 4  is H or a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or a combination thereof,  
 further wherein said compound inhibits mitochondrial fusion.  
 
     
     
         69 . The compound of  claim 68 , wherein W is N and R 1  is H.  
     
     
         70 . The compound of  claim 69 , wherein R 3  and R 4  are O.  
     
     
         71 . The compound of  claim 70 , wherein the compound is 2-((E-((E)-2-(2-chlorophenylimino)-4-oxothiazolin-5-ylidene)methyl)-6-ethoxyphenolate.  
     
     
         72 . The compound of  claim 68 , wherein W is S, and R1 is a substituted piperidine.  
     
     
         73 . The compound of  claim 68 , wherein the compound is (E)-5-(4-methylbenzylidene)-3-((4-methylpiperidin-1-yl)methyl)-2-thioxothiazolidin-4-one.  
     
     
         74 . A pharmaceutical composition comprising the compound of any one of claims  68 - 73  and a pharmaceutically acceptable carrier.  
     
     
         75 . A method of modulating mitochondrial fusion in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  68 - 73 .  
     
     
         76 . A method of increasing apoptosis in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  68 - 73 .  
     
     
         77 . A method of treating a condition associated with apoptosis, wherein the treatment includes increasing apoptosis comprising administering to a subject in need thereof a compound of any one of claims  68 - 73 .  
     
     
         78 . The method of  claim 77 , wherein the condition is neoplasia.  
     
     
         79 . A compound selected from the group consisting of (Z)-ethyl-5-benzylidene-2-(4-nitrophenylamino)-4-oxo-tetrahydrothiophene-3-carboxylate, 5-(5-chloro-2(2-(m-tolyloxy)ethoxy)benzylidene)-2-thioxo-dihydropyrimidin-4,6(1H,5H)-dione, and 1-(3-bromophenyl)-3-(ethoxycarbonyl)-2-methyl-1H-benzo[g]indol-5-olate, wherein said compound inhibits mitochondrial fusion.  
     
     
         80 . A pharmaceutical composition comprising the compound of  claim 79  and a pharmaceutically acceptable carrier.  
     
     
         81 . A method of modulating mitochondrial fusion in a cell comprising contacting the cell with a formulation comprising the compound of  claim 79 .  
     
     
         82 . A method of increasing apoptosis in a cell comprising contacting the cell with a formulation comprising the compound of  claim 79 .  
     
     
         83 . A method of treating a condition associated with apoptosis, wherein the treatment includes increasing apoptosis comprising administering to a subject in need thereof a compound of  claim 79 .  
     
     
         84 . The method of  claim 83 , wherein the condition is neoplasia.  
     
     
         85 . A method for treating a condition associated with apoptosis comprising administering to a subject in need of such treatment an agent capable of regulating mitochondrial fission.  
     
     
         86 . A method for treating a condition associated with apoptosis comprising administering to a subject in need of such treatment an agent capable of regulating mitochondrial fusion.  
     
     
         87 . A method of treating a condition associated with apoptosis, wherein the treatment includes increasing apoptosis comprising administering to a subject in need of such treatment an agent capable of increasing mitochondrial fission or decreasing mitochondrial fusion.  
     
     
         88 . A method of treating a condition associated with apoptosis, wherein the treatment includes decreasing apoptosis comprising administering to a subject in need of such treatment an agent capable of decreasing mitochondrial fission or increasing mitochondrial fusion.  
     
     
         89 . A method of screening for an agent capable of decreasing mitochondrial fission comprising identifying an agent that suppresses the growth defect of a cell with decreased mitochondrial fusion.  
     
     
         90 . An agent identified by the method of  claim 89 .  
     
     
         91 . A method of screening for an agent capable of increasing mitochondrial fission comprising identifying an agent that suppresses the growth on a non-fermentable carbon source of a cell in wild type and does not suppress the growth on a non-fermentable carbon source of the cell defective in mitochondrial fission.  
     
     
         92 . A compound identified by the method of  claim 91 .  
     
     
         93 . A compound of the general formula:  
       
         
           
           
               
               
           
         
       
       wherein W is OH or an ester, wherein said ester may contain a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; 
 R 1  is independently H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkenyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C1-C18 alkynl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; a C3-C18 aryl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a C5-C18 cycloalkyl which may contain a side group, may contain a bridge, may contain a heteroatom or may be substituted, or combinations thereof; or a halogen; 
 R 2  is H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof; or a halogen;  
 R 3  is H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof;  
 R4 is H; a C1-C18 alkyl, which may be branched, may contain a heteroatom or may be substituted, or combinations thereof;  
 further wherein said compound causes the formation of net like structures in mitochondria.  
 
 
     
     
         94 . The compound of  claim 93 , wherein R1 is a halogen.  
     
     
         95 . The compound of  claim 94 , wherein W is OH.  
     
     
         96 . The compound of  claim 95 , wherein R4 is a carbonyl radical.  
     
     
         97 . The compound of  claim 96 , wherein the compound is 6-chloro-4-hydroxy-8-methylquinoline-3-carboxylic acid.  
     
     
         98 . The compound of  claim 93 , wherein R2 is a halogen.  
     
     
         99 . The compound of  claim 98 , wherein W is an ester.  
     
     
         100 . The compound of  claim 99 , wherein the compound is 7-chloro-4-(4-chloro-3,5-dimethylphenoxy)quinolone.  
     
     
         101 . A pharmaceutical composition comprising the compound of any one of claims  93 -100 and a pharmaceutically acceptable carrier.  
     
     
         102 . A method of modulating mitochondrial fission in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  93 - 100 .  
     
     
         103 . A method of inhibiting apoptosis in a cell comprising contacting the cell with a formulation comprising the compound of any one of claims  93 - 100 .  
     
     
         104 . A method of treating a condition associated with apoptosis, wherein the treatment includes decreasing apoptosis comprising administering to a subject in need thereof a compound of any one of claims  93 - 100 .  
     
     
         105 . The method of  claim 104 , wherein the condition is selected from the group consisting of Parkinson's disease, ALS, stroke, heart attack, congestive heart failure, transplantation, alcoholic hepatitis, and drug-induced liver toxicity.