Fluorescent dyes (AIDA) for solid phase and solution phase screening

Abstract

The invention relates to new fluorescent dyes of formula (I) which can be used in high throughput screening both, on the solid phase as well as in homogeneous solution.

1 - 11 . (canceled)  
   
   
       12 . Compounds comprising structures of the formulae II-III  
       A-B-D-C-D′-   (Formula (II))  A-B-D- and -D′-C   (Formula (III))  
     wherein 
 A is selected from the group consisting of functionalized polystyrene based resins, polyacrylamide based polymers, polystyrene/polydimethylacrylamide composites, PEGA resins, polystyrene-polyoxyethylene based supports, Tentagel resins, PEG-polystyrene graft polymeric supports, glass surfaces, functionalized surfaces, materials grafted with functionalized surfaces, and polyethylenglycol;  
 B is a linker allowing cleavage of fluorescent conjugates of formula (II-III) for liberation of the D and C containing fragments;  
 C is a compound selected from formula (I)  
                     
 wherein  
 one of the radicals R 1  or R 2  and one of the radicals R 3  or R 4  is hydrogen and the other is independently —COOH, —COOR 7 , —CONH 2 , —CONH(CH 2 ) n OH, —CONR 8 R 9 , —CH 2 OH, —CH 2 NH 2 , —NO 2 , —NR 10 R 11 , —NHCOR 12 , Cl, Br, F, —CF 3 , —N═C═O, —N═C═S, —SO 3 H, —SO 2 NH(CH 2 ) n NH 2 , (C 1 -C 4 ) alkyl, (C 1 -C 16 )-alkyl substituted at the terminal carbon with —COOH, —COOR 7 , —CONH 2 , —CONR 8 R 9 , —CONH(CH 2 ) n OH, —CH 2 OH, —CH 2 NH 2 , —N═C═O, —N═C═S, —SO 3 H, —SO 2 NH(CH 2 ) n NH 2 , —CONH(CH 2 ) n NH 2 , and the —NH 2  group could also be substituted by (C 1 -C 4 ) alkyl or a commonly used amino protecting group;  
 and one of the radicals R 5  or R 6  is hydrogen and the other is hydrogen, halogen, —NO 2 , —NR 10 R 11 , —NHCOR 12 , (C 1 -C 4 ) alkyl, (C 1 -C 16 )-alkyl substituted at the terminal carbon with —COOH, —COOR 7 , —CONH 2 , —CONR 8 R 9 , —CONH(CH 2 ) n OH, —CH 2 OH, —CH 2 NH 2 , —N═C═O, —N═C═S, —SO 3 H, —SO 2 NH(CH 2 ) n NH 2 , —CONH(CH 2 ) n NH 2 , wherein and the —NH 2  group could also be substituted by (C 1 -C 4 ) alkyl or a commonly used amino protecting group;  
 n is 2-8;  
 with the proviso that only one of R 1 -R 6  is nitro;  
 R 7  is a commonly used carboxyl protecting or carboxyl activating group;  
 R 8  or R 9  is hydrogen and the other is lower alkyl (C 1 -C 4 ), phenyl, benzyl, or R 8  and R 9  are part of a 5 or 6 membered ring;  
 R 10  and R 11  are independently hydrogen or (C 1 -C 4 )alkyl; and  
 R 12  is (C 1 -C 10 )alkyl, phenyl, which both can be substituted by (C 1 -C 4 ) alkyl, protected amino group or halogen; and  
 D and D′ are independently a bond or a spacer selected from α,ω-diamino-alkanes, diaminocyclohexyl, bis-(aminomethyl)-substituted phenyl, α-amino-ω-hydroxy-alkanes, alkylamines, cyclic alkylamines or cyclic alkyldiamines or amino acids without or with additional functionality in the side chain.  
 
   
   
       13 . Compounds of  claim 12 , wherein 
 B is selected from benzyl, benzhydryl, benzhydryliden, trityl, xanthenyl, benzoin, silicon, or allyl based linkers.    
   
   
       14 . Compounds of  claim 12  of the following structures:  
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
   
   
       15 . Compounds of  claim 12  wherein the amino protecting group is tert-butyloxycarbonyl, 9-fluorenylmethoxycarbonyl, phthalimido, trifluoroacetamido, methoxycarbonyl, ethoxycarbonyl, benzyloxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, or 2-(trimethylsilyl)ethoxy-carbonyl.  
   
   
       16 . Compounds of  claim 12  wherein C is of the following structures: