Inhibitors of the Interaction between p53 and MDM2

Abstract

The present invention relates to compounds capable of binding to the oncogene protein MDM2, processes for the preparation of such compounds, pharmaceutical preparations comprising such compounds, and uses of said compounds, e.g. in the therapeutic (including prophylactic) treatment of an animal or especially of the human body. The present further relates to methods of and compounds for inhibiting the growth of tumor cells which comprise the wild type p53 suppressor by interfering with the interaction between human p53 and human MDM2.

1 - 23 . (canceled)  
     
     
         24 . A peptide which binds to a DM2 protein and inhibits binding of said DM2 protein to a p53 protein, the peptide being a derivative of an amino acid sequence that comprises an amino acid motif comprising at least 8 consecutive amino acids of the formula  
       
         
           
                 
                 
                 
                 
               
                     
                 
                   R 1 -X 1 -F-X 2 -R 2 -R 3 -W-X 3 -X 4 -R 4   
                   (I), 
                   (SEQ ID NO:4) 
                     
                 
                     
                 
             
                
                
                
               
            
           
         
         wherein  
         R 1  is proline (P), leucine (L), glutamic acid (E), cysteine (C) or glutamine (Q),  
         X stands for any natural amino acid,  
         R 2  is arginine (R), histidine (H), glutamic acid, cysteine, serine, or aspartic acid (D),  
         R 3  is histidine (H), phenylalanine (F) or tyrosine (Y),  
         R 4  is phenylalanine (F), glutamine (Q) or leucine (L), and  
         F is phenylalanine and W is tryptophan,  
         said derivative being modified from said amino acid sequence by substitution, whereby one or more amino acids in said amino acid motif are each replaced by a non-natural amino acid.  
       
     
     
         25 . A peptide according to  claim 24  wherein said amino acid motif comprises at least the eight consecutive amino acids  
       
         
           
                 
                 
                 
                 
                 
               
                     
                     
                 
                     
                   F-X 2 -R 2 -R 3 -W-X 3 -X 4 -R 4   
                   (Ib) 
                   (SEQ ID NO:10) 
                     
                 
                     
                     
                 
             
                
                
                
               
            
           
         
       
       from the sequence of formula (I) (SEQ ID NO:4),  
     
     
         26 . A peptide according to  claim 24 , wherein the MDM2 is human DM2.  
     
     
         27 . A peptide according to  claim 24 , wherein the p53 is human p53.  
     
     
         28 . A peptide according to  claim 24 , wherein the modification is such that an α-helix conformation in the peptide is induced, increased or maintained.  
     
     
         29 . A peptide according to  claim 24 , wherein R 2 , X 3  and/or X 4  are, independently from one another, replaced by α,α-disubstituted amino acid residue, α-aminoisobutyric acid, 1-aminocyclopropane-1-carboxylic acid, 1-amino-cyclopentane-1-carboxylic acid, 1-aminocyclohexane-1-carboxylic acid, 4-amino piperidine-4-carboxylic acid, or 1-aminocycloheptane-1-carboxylic acid.  
     
     
         30 . A peptide according to  claim 29 , wherein R 2  is replaced by α-aminoisobutyric acid (Aib), X 3  is replaced by α-aminoisobutyric acid and/or X 4  is replaced by 1-aminocyclopropane-1-carboxylic acid (Ac 3 c).  
     
     
         31 . A peptide according to  claim 24 , wherein the modification includes replacement of an amino acid with a non-natural amino acid mimetic thereof.  
     
     
         32 . A peptide according to  claim 30 , wherein the modification further includes replacement of an amino acid with a non-natural amino acid mimetic thereof.  
     
     
         33 . A peptide according to  claim 24 , consisting of no more than fifteen amino acids.  
     
     
         34 . A peptide according to  claim 25 , which is a said derivative of an amino acid sequence that consists of an amino acid motif of the formula  
       
         
           
                 
                 
                 
                 
                 
               
                     
                     
                 
                     
                   F-X 2 -R 2 -R 3 -W-X 3 -X 4 -R 4   
                   (Ib) 
                   (SEQ ID NO:10) 
                     
                 
                     
                     
                 
             
                
                
                
               
            
           
         
         wherein  
         X 2  is methionine, isoleucine, threonine, arginine, alanine or serine;  
         X 3  is glutamic acid, threonine, alanine, phenylalanine or serine; and  
         X 4  is glycine, glutamine, threonine, alanine or aspartic acid.  
       
     
     
         35 . A peptide according to  claim 34 , wherein 
 X 2  is methionine;    X 3  is glutamic acid; and/or    X 4  is glycine.    
     
     
         36 . A peptide according to  claim 25 , which is a said derivative of an amino acid sequence that consists of an amino acid motif of the formula  
       
         
           
                 
                 
                 
                 
               
                     
                 
                   X 1 -F-X 2 -R 2 -R 3 -W-X 3 -X 4 -R 4   
                   (Ic) 
                   (SEQ ID NO:11) 
                     
                 
                     
                 
             
                
                
                
               
            
           
         
         wherein  
         X 1  is arginine, asparagine, alanine, threonine or valine;  
         X 2  is methionine, isoleucine, threonine, arginine, alanine or serine;  
         X 3  is glutamic acid, threonine, alanine, phenylalanine or serine; and  
         X 4  is glycine, glutamine, threonine, alanine or aspartic acid.  
       
     
     
         37 . A peptide according to  claim 36 , wherein 
 X 1  is arginine;    X 2  is methionine;    X 3  is glutamic acid; and/or    X 4  is glycine.    
     
     
         38 . A peptide according to  claim 24  carrying one or more protecting groups.  
     
     
         39 . A peptide according to  claim 24  which is in the form of a salt.  
     
     
         40 . A peptide according to  claim 24  which is in cyclic form.  
     
     
         41 . A peptide according to  claim 40 , which contains a disulphide bridge, a thioether bridge or a lactam.  
     
     
         42 . A conjugate comprising the peptide of  claim 24  covalently attached to another peptide or protein.  
     
     
         43 . A conjugate comprising a peptide of  claim 24 , labeled directly or by way of a spacer or linker group with an enzyme, a fluorescent marker, a chemiluminescent marker, a metal chelate, paramagnetic particles or biotin.  
     
     
         44 . A method for inhibiting the binding of a DM2 protein to a p53 protein comprising contacting said DM2 protein with a peptide according to  claim 24 .  
     
     
         45 . A composition comprising a peptide according to  claim 24 , in admixture with at least one pharmaceutically acceptable carrier.  
     
     
         46 . A peptide which binds to a DM2 protein and inhibits binding of said DM2 protein to a p53 protein, the peptide consisting of no more than 15 amino acids and being a derivative of an amino acid sequence that comprises an amino acid motif comprising at least the eight consecutive amino acids  
       
         
           
                 
                 
                 
                 
               
                     
                     
                 
                     
                   F-X 2 -R 2 -R 3 -W-X 3 -X 4 -R 4  (Ib) 
                   (SEQ ID NO:10) 
                     
                 
                     
                     
                 
             
                
                
                
               
            
           
         
       
       from the sequence  
       
         
           
                 
                 
                 
                 
               
                     
                     
                 
                     
                   R 1 -X 1 -F-X 2 -R 2 -R 3 -W-X 3 -X 4 -R 4  (I), 
                   (SEQ ID NO:4) 
                     
                 
                     
                     
                 
             
                
                
                
               
            
           
         
         wherein  
         R 1  is proline (P), leucine (L), glutamic acid (E), cysteine (C) or glutamine (Q),  
         X 1  is any natural amino acid,  
         X 2  is methionine (M),  
         R 2  is arginine (R), histidine (H), glutamic acid (E), cysteine (C), serine (S), or aspartic acid (D),  
         R 3  is phenylalanine (F) or tyrosine (Y),  
         X 3  is glutamic acid (E),  
         X 4  is any natural amino acid  
         R 4  is phenylalanine (F), glutamine (Q) or leucine (L), and  
         F is phenylalanine and W is tryptophan,  
         said derivative being modified from said amino acid sequence by substitution, whereby a plurality of amino acids in said amino acid motif are each replaced by a nonnatural amino acid, wherein said modification includes:  
         (1) R 2  being replaced by α-aminoisobutyric acid (Aib), X 3  being replaced by α-aminoisobutyric acid and/or X 4  being replaced by 1-amino-cyclopropane-1-carboxylic acid (Ac 3 c); and  
         (2) replacement of a plurality of amino acids with a non-natural amino acid mimetic thereof, said replacement including R 3  being replaced by a structurally related analogue.  
       
     
     
         47 . A conjugate comprising the peptide of  claim 46  covalently attached to another peptide or protein.  
     
     
         48 . A conjugate comprising a peptide of  claim 46 , labeled directly or by way of a spacer or linker group with an enzyme, a fluorescent marker, a chemiluminescent marker, a metal chelate, paramagnetic particles or biotin.  
     
     
         49 . A method for inhibiting the binding of a DM2 protein to a p53 protein comprising contacting said DM2 protein with a peptide according to  claim 46 .  
     
     
         50 . A composition comprising a peptide according to  claim 46 , in admixture with at least one pharmaceutically acceptable carrier.