Small-molecule inhibitors targeting g-protein-coupled rho guanine nucleotide exchange factors

Abstract

Provided are inhibitors of Rho GTPase activation, and, in particular, compounds that inhibit RhoA activation by an RhoGEF. Also provided are related pharmaceutical compositions and methods. Also provided are methods of inhibiting Rho GTPase activation. Also provided are methods of screening for compounds that inhibit Rho GTPase activation by a RhoGEF.

1 . A compound having the structure of Formula I: 
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt thereof or tautomer thereof, 
         wherein: 
         R 1  is aryl, heteroaryl, or heterocyclyl, each optionally substituted with one or more R 1A  with the proviso that R 1  is not furanyl or furanyl substituted with optionally substituted aryl;
 each R 1A  is independently selected from the group consisting of hydroxy, halo, cyano, nitro, —SO 2 OH, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, isocyanato, thiocyanato, isothiocyanato, amino, optionally substituted C 3-7  cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, C 1-6  alkyl optionally substituted with up to 5 R 1B , and C 1-6  alkoxy optionally substituted with up to 5 R 1B ;
 each R 1B  is independently selected from the group consisting of OR 1C , C-carboxy, O-carboxy, aryl, and heteroaryl, said aryl or heteroaryl each optionally substituted with one or more R 1D ; 
 each R 1C  is independently selected from the group consisting of aryl, and heteroaryl, said aryl or heteroaryl each optionally substituted with one or more R 1E ; 
 each R 1D  is independently selected from the group consisting of hydroxy, halo, cyano, nitro, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, isocyanato, thiocyanato, isothiocyanato, amino, optionally substituted C 3-7  cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, C 1-6  alkyl optionally substituted with up to 5 fluoro, and C 1-6  alkoxy optionally substituted with up to 5 fluoro;
 each R 1E  is independently selected from the group consisting of hydroxy, halo, cyano, nitro, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, isocyanato, thiocyanato, isothiocyanato, amino, optionally substituted C 3-7  cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, C 1-6  alkyl optionally substituted with up to 5 fluoro, and C 1-6  alkoxy optionally substituted with up to 5 fluoro; 
 
 
 
         R 2  is H (hydrogen), aryl, or heteroaryl, said aryl or heteroaryl each optionally substituted with one or more R 2A ;
 each R 2A  is independently selected from the group consisting of hydroxy, halo, cyano, nitro, O-carbamyl, N-carbamyl, C-amido, N-amido, C-carboxy, O-carboxy, amino, optionally substituted C 3-7  cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, C 1-6  alkyl optionally substituted with up to 5 fluoro, and C 1-6  alkoxy optionally substituted with up to 5 fluoro; 
 
         R 3  is H (hydrogen), aryl, or heteroaryl, said aryl or heteroaryl each optionally substituted with one or more R 3A , provided that one of R 2  and R 3  is H (hydrogen) and one of R 2  and R 3  is not H (hydrogen);
 each R 3A  is independently selected from the group consisting of hydroxy, halo, cyano, nitro, O-carbamyl, N-carbamyl, C-amido, N-amido, C-carboxy, O-carboxy, amino, optionally substituted C 3-7  cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, C 1-6  alkyl optionally substituted with up to 5 fluoro, and C 1-6  alkoxy optionally substituted with up to 5 fluoro; 
 R 4  is H (hydrogen), or C 1-6  alkyl; and 
 R 5  is H (hydrogen), or C 1-6  alkyl, or optionally R 4  and R 5  together are oxo, with the proviso that the compound is not: 
 
       
       
         
           
           
               
               
           
         
         4-(4-Iodo-benzylidene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione,
 4-(Dimethyl-furan-2-ylmethylene)-1-(3-trifluoromethyl-phenyl)-pyrazolidine-3,5-dione, 
 Acetic acid 5-[1-(4-iodo-phenyl)-3,5-dioxo-pyrazolidin-4-ylidenemethyl]-furan-2-ylmethyl ester, 
 4-(4-Bromo-furan-2-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-(5-methyl-furan-2-ylmethylene)-pyrazolidine-3,5-dione, 
 4-(5-Bromo-furan-2-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-[5-(2-methoxy-phenyl)-thiophen-2-ylmethylene]-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-thiophen-3-ylmethylene-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-(5-phenyl-thiophen-2-ylmethylene)-pyrazolidine-3,5-dione, 
 4-(5-Methyl-furan-2-ylmethylene)-1-(3-trifluoromethyl-phenyl)-pyrazolidine-3,5-dione, 
 4-Thiophen-2-ylmethylene-1-(3-trifluoromethyl-phenyl)-pyrazolidine-3,5-dione, 
 4-(4-Bromo-furan-2-ylmethylene)-1-(3-trifluoromethyl-phenyl)-pyrazolidine-3,5-dione, 
 Acetic acid 5-[3,5-dioxo-1-(3-trifluoromethyl-phenyl)-pyrazolidin-4-ylidenemethyl]-furan-2-ylmethyl ester, 
 4-(7-Bromo-8-hydroxy-quinolin-5-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-[5-(4-nitro-phenyl)-furan-2-ylmethylene]-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-[5-(2-nitro-phenyl)-furan-2-ylmethylene]-pyrazolidine-3,5-dione, 
 4-(5-Hydroxymethyl-furan-2-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-(5-nitro-thiophen-2-ylmethylene)-pyrazolidine-3,5-dione, 
 4-(5-Hydroxymethyl-furan-2-ylmethylene)-1-(3-trifluoromethyl-phenyl)-pyrazolidine-3,5-dione, 
 4-(5-Nitro-thiophen-2-ylmethylene)-1-(3-trifluoromethyl-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-(5-nitro-furan-2-ylmethylene)-pyrazolidine-3,5-dione, 
 4-(4, 5-Dimethyl-furan-2-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 5-[1-(4-Iodo-phenyl)-3,5-dioxo-pyrazolidin-4-ylidenemethyl]-furan-2-sulfonic acid, 
 4-[1-(4-Iodo-phenyl)-3,5-dioxo-pyrazolidin-4-ylidenemethyl]-benzonitrile, 
 1-(4-Iodo-phenyl)-4-(3-methyl-thiophen-2-ylmethylene)-pyrazolidine-3,5-dione, 
 4-(4-Bromo-thiophen-2-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-(5-methyl-thiophen-2-ylmethylene)-pyrazolidine-3,5-dione, 
 4-Benzo[b]thiophen-2-ylmethylene-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-(4-nitro-thiophen-2-ylmethylene)-pyrazolidine-3,5-dione, 
 5-[1-(4-Iodo-phenyl)-3,5-dioxo-pyrazolidin-4-ylidenemethyl]-thiophene-3-carboxylic acid, 
 4-[1-(4-Iodo-phenyl)-3,5-dioxo-pyrazolidin-4-ylidenemethyl]-thiophene-3-carboxylic acid, 
 2-[3,5-Dioxo-1-(3-trifluoromethyl-phenyl)-pyrazolidin-4-ylidenemethyl]-thiophene-3-carboxylic acid, 
 4-(4-Nitro-thiophen-2-ylmethylene)-1-(3-trifluoromethyl-phenyl)-pyrazolidine-3,5-dione, 
 4-[3,5-Dioxo-1-(3-trifluoromethyl-phenyl)-pyrazolidin-4-ylidenemethyl]-thiophene-3-carboxylic acid, 
 {3-[3,5-Dioxo-1-(3-trifluoromethyl-phenyl)-pyrazolidin-4-ylidenemethyl]-thiophen-2-ylsulfanyl-acetic acid, 
 2-[1-(4-Iodo-phenyl)-3,5-dioxo-pyrazolidin-4-ylidenemethyl]-thiophene-3-carboxylic acid, 
 4-(4-Hydroxymethyl-furan-3-ylmethylene)-1-(3-trifluoromethyl-phenyl)-pyrazolidine-3,5-dione, 
 4-benzo[b]thiophen-2-ylmethylene-1-(3-trifluoromethyl-phenyl)-pyrazolidine-3,5-dione, 
 5-[3,5-Dioxo-1-(3-trifluoromethyl-phenyl)-pyrazolidin-4-ylidenemethyl]-thiophene-3-carboxylic acid, 
 5-[1-(4-Iodo-phenyl)-3,5-dioxo-pyrazolidin-4-ylidenemethyl]-furan-2-carboxylic acid, 
 5-(1-Biphenyl-4-yl-3,5-dioxo-pyrazolidin-4-ylidenemethyl)-furan-2-carboxylic acid, 
 4-(4-Hydroxy-naphthalen-1-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-(1-oxy-pyridin-4-ylmethylene)-pyrazolidine-3,5-dione, 
 4-(1H-Indol-5-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 4-(4-Hydroxymethyl-furan-3-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodophenyl)-4-(1H-pyrrol-2-ylmethylene)-pyrazolidine-3,5-dione, 
 1-(4-Iodophenyl)-4-(1-methyl-1H-pyrrol-2-ylmethylene)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-[1-(2-nitro-benzyl)-1H-pyrrol-2-ylmethylene]-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl-4-(1H-pyrazol-3-ylmethylene)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl-4-(1H-pyrazol-3-ylmethylene)-pyrazolidine-3,5-dione, 
 4-(2, 3-Dihydro-benzofuran-5-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-quinolin-2-ylmethylene-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-quinolin-4-ylmethylene-pyrazolidine-3,5-dione, 
 4-[3-(2-Hydroxy-ethoxy)-benzylidene]-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-(5-naphthalen-2-yl-thiophen-2-ylmethylene)-pyrazolidine-3,5-dione, 
 3-{5-[1-(4-Iodo-phenyl)-3,5-dioxo-pyrazolidin-4-ylidenemethyl]-furan-2-yl}-thiophene-2-carboxylic acid methyl ester, 
 4-(5-Chloro-thiophen-2-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 [1-(4-Iodo-phenyl)-4-thiazol-2-ylmethylene-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-(1-methyl-1H-imidazol-2-ylmethylene)-pyrazolidine-3,5-dione, 
 4-(2-Diethylamino-thiazol-5-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 4-[2-(4-Benzyl-piperazin-1-yl)-thiazol-5-ylmethylene]-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 1-(4-Iodo-phenyl)-4-[2-(4-methoxy-phenoxy)-thiazol-5-ylmethylene]-pyrazolidine-3,5-dione, 
 4-(9-Ethyl-9H-carbazol-3-ylmethylene)-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, or 4-Benzo[b]thiophen-3-ylmethylene-1-(4-iodo-phenyl)-pyrazolidine-3,5-dione, 
 or a pharmaceutically acceptable salt thereof or tautomer thereof. 
 
       
     
     
         2 . The compound of  claim 1 , wherein R 2  is H (hydrogen), or wherein R 3  is H (hydrogen). 
     
     
         3 . (canceled) 
     
     
         4 . The compound of  claim 1 , wherein R 2  is phenyl optionally substituted with one or more R 2A ; and
 each R 2A  is independently selected from the group consisting of halo, C 1-6  alkyl optionally substituted with up to 5 fluoro, and C 1-6  alkoxy optionally substituted with up to 5 fluoro.   
     
     
         5 . The compound of  claim 4 , wherein R 2  is 
       
         
           
           
               
               
           
         
       
     
     
         6 . The compound of  claim 5 , wherein R 2A  is fluoro, chloro, bromo, iodo, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, or trifluoromethoxy. 
     
     
         7 . The compound of  claim 1 , wherein R 3  is phenyl optionally substituted with one or more R 3A ; and
 each R 3A  is independently selected from the group consisting of halo, C 1-6  alkyl optionally substituted with up to 5 fluoro, and C 1-6  alkoxy optionally substituted with up to 5 fluoro.   
     
     
         8 . The compound of  claim 7 , wherein R 3  is 
       
         
           
           
               
               
           
         
       
     
     
         9 . The compound of  claim 8 , wherein R 3A  is fluoro, chloro, bromo, iodo, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, or trifluoromethoxy. 
     
     
         10 . The compound of  claim 1 , wherein R 1  is heteroaryl, phenyl or heterocyclyl, each optionally substituted with one or more R 1A ;
 each R 1A  is independently selected from the group consisting of hydroxy, fluoro, chloro, bromo, halo, cyano, amino, C 1-3  alkyl, C 1-3  alkoxy, C 1-6  alkyl, C 1-6  alkyl substituted with R 1B , C 1-4  alkoxy, and C 1-4  alkoxy substituted with R 1B ;
 each R 1B  is independently selected from the group consisting of —OR 1C , phenyl, and heteroaryl, said phenyl optionally substituted with one or more R 1D ; 
 each R 1C  is independently selected from the group consisting of phenyl, and heteroaryl, said phenyl or heteroaryl each optionally substituted with one or more R 1E ; 
 each R 1D  is independently selected from the group consisting of hydroxy, fluoro, chloro, bromo, halo, cyano, amino, C 1-3  alkyl optionally substituted with up to 5 fluoro, and C 1-3  alkoxy optionally substituted with up to 5 fluoro; and, 
 each R 1E  is independently selected from the group consisting of hydroxy, fluoro, chloro, bromo, halo, cyano, amino, C 1-3  alkyl optionally substituted with up to 5 fluoro, and C 1-3  alkoxy optionally substituted with up to 5 fluoro. 
   
     
     
         11 .- 13 . (canceled) 
     
     
         14 . The compound of  claim 1 , wherein R 1A  is C 1-4  alkoxy substituted with R 1B ; and R 1B  is phenyl optionally substituted with one or more R 1D . 
     
     
         15 . The compound of  claim 10 , wherein R 1  is 1H-isoindolyl. 
     
     
         16 . The compound of  claim 1 , wherein R 4  is H (hydrogen); and R 5  is H (hydrogen). 
     
     
         17 . The compound of  claim 1 , wherein R 4  and R 5  together are oxo. 
     
     
         18 . The compound of  claim 1 , wherein the compound having the structure of Formula I has the structure of Formula Ia, Ib, or Ib: 
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt thereof. 
       
     
     
         19 . The compound of  claim 1 ,
 wherein:   R 1  is   
       
         
           
           
               
               
           
         
         each R 6  is independently selected from the group consisting of H (hydrogen), hydroxy, halo, C 1-3  alkyl, C 1-3  alkoxy, and C 1-4  alkoxy substituted with R 1B ;
 each R 1B  is independently selected from the group consisting of —OR 1C , and phenyl, said phenyl optionally substituted with one or more R 1D ; 
 each R 1C  is phenyl optionally substituted with one or more R 1E ; 
 each R 1D  is independently selected from the group consisting of fluoro, chloro, bromo, cyano, C 1-3  alkyl optionally substituted with up to 5 fluoro, and C 1-3  alkoxy optionally substituted with up to 5 fluoro; and 
 
         each R 1E  is independently selected from the group consisting of fluoro, chloro, bromo, cyano, C 1-3  alkyl optionally substituted with up to 5 fluoro, and C 1-3  alkoxy optionally substituted with up to 5 fluoro. 
       
     
     
         20 . The compound of  claim 19 ,
 wherein:   R 1  is   
       
         
           
           
               
               
           
         
          or; 
         each R 6  is independently selected from the group consisting of hydroxy, fluoro, chloro, bromo, methyl, methoxy, ethoxy, and C 1-3  alkoxy substituted with R 1B ;
 each R 1B  is independently selected from the group consisting of —OR 1C , and phenyl, said phenyl optionally substituted with one or more R 1D ; 
 each R 1C  is phenyl optionally substituted with one or more R 1E ; 
 each R 1D  is independently selected from the group consisting of fluoro, chloro, bromo, cyano, methyl, ethyl, methoxy, and ethoxy; and 
 each R 1E  is independently selected from the group consisting of fluoro, chloro, bromo, cyano, methyl, ethyl, methoxy, and ethoxy. 
 
       
     
     
         21 . The compound of  claim 1  having the structure: 
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt thereof. 
       
     
     
         22 . (canceled) 
     
     
         23 . A composition comprising a pharmaceutically acceptable excipient, and a compound of  claim 1 , or a pharmaceutically acceptable salt thereof or tautomer thereof. 
     
     
         24 .- 58 . (canceled) 
     
     
         59 . A method of inhibiting Rho GTPase activation comprising contacting a RhoGEF with a compound having the structure of Formula I: 
       
         
           
           
               
               
           
         
         or a pharmaceutically acceptable salt thereof, 
         wherein: 
         R 1  is aryl, heteroaryl, or heterocyclyl, each optionally substituted with one or more R 1A ;
 each R 1A  is independently selected from the group consisting of hydroxy, halo, cyano, nitro, —SO 2 OH, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, isocyanato, thiocyanato, isothiocyanato, amino, optionally substituted C 3-7  cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, C 1-6  alkyl optionally substituted with up to 5 R 1B , and C 1-6  alkoxy optionally substituted with up to 5 R 1B ;
 each R 1B  is independently selected from the group consisting of OR 1C , aryl, and heteroaryl, said aryl or heteroaryl each optionally substituted with one or more R 1D ; 
 each R 1C  is independently selected from the group consisting of aryl, and heteroaryl, said aryl or heteroaryl each optionally substituted with one or more R 1E ; 
 each R 1D  is independently selected from the group consisting of hydroxy, halo, cyano, nitro, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, isocyanato, thiocyanato, isothiocyanato, amino, optionally substituted C 3-7  cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, C 1-6  alkyl optionally substituted with up to 5 fluoro, and C 1-6  alkoxy optionally substituted with up to 5 fluoro;
 each R 1E  is independently selected from the group consisting of hydroxy, halo, cyano, nitro, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, O-carboxy, isocyanato, thiocyanato, isothiocyanato, amino, optionally substituted C 3-7  cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, C 1-6  alkyl optionally substituted with up to 5 fluoro, and C 1-6  alkoxy optionally substituted with up to 5 fluoro; 
 
 
 
         R 2  is H (hydrogen), aryl, or heteroaryl, said aryl or heteroaryl each optionally substituted with one or more R 2A ;
 each R 2A  is independently selected from the group consisting of hydroxy, halo, cyano, nitro, O-carbamyl, N-carbamyl, C-amido, N-amido, C-carboxy, O-carboxy, amino, optionally substituted C 3-7  cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, C 1-6  alkyl optionally substituted with up to 5 fluoro, and C 1-6  alkoxy optionally substituted with up to 5 fluoro; 
 
         R 3  is H (hydrogen), aryl, or heteroaryl, said aryl or heteroaryl each optionally substituted with one or more R 3A , provided that one of R 2  and R 3  is H (hydrogen) and one of R 2  and R 3  is not H (hydrogen);
 each R 3A  is independently selected from the group consisting of hydroxy, halo, cyano, nitro, O-carbamyl, N-carbamyl, C-amido, N-amido, C-carboxy, O-carboxy, amino, optionally substituted C 3-7  cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclyl, C 1-6  alkyl optionally substituted with up to 5 fluoro, and C 1-6  alkoxy optionally substituted with up to 5 fluoro; 
 
         R 4  is H (hydrogen), or C 1-6  alkyl; and 
         R 5  is H (hydrogen), or C 1-6  alkyl, or optionally R 4  and R 5  together are oxo. 
       
     
     
         60 .- 68 . (canceled) 
     
     
         69 . The method of  claim 59 , wherein a therapeutically effective amount of the compound having the structure of Formula I is administered to a subject in need of treatment for breast cancer, leukemia or lung cancer.