Methods for designing inhibitors of serine/threonine kinases and tyrosine kinases

Abstract

The invention relates to methods for designing inhibitors of serine/threonine kinases and tyrosine kinases, particularly MAP kinases, through the use of ATP-binding site mutants of those kinases. The methods of this invention take advantage of the fact that the mutant kinases are capable of binding inhibitory compounds of other kinases with greater affinity than the corresponding wild-type kinase. The invention further relates to the mutant kinases themselves and crystallizable co-complexes of the mutant kinase and the inhibitory compound.

1 - 9 . (cancelled).  
     
     
         10 . A mutant second serine/threonine kinase or tyrosine kinase characterized by: 
 a. at least one amino acid substitution in an ATP binding site as compared to a corresponding naturally occurring second kinase;    b. the ability to bind a K i  or a K d  of less than 10 μM a compound that binds to an ATP binding site of a first serine/threonine kinase or tyrosine kinase; and    c. the ability to bind said compound with at least a 10-fold lower K i  or K d  than the K i  or K d  for said compound with said second kinase.    
     
     
         11 . The mutant second kinase according to  claim 10 , wherein said first and said second kinases are MAP kinases.  
     
     
         12 . The mutant second kinase according to  claim 11 , wherein said mutant second kinase is selected from: 
 a. a mutant ERK2 consisting of the amino acid sequence as set forth in SEQ ID NO:2, wherein amino acid 105 is threonine or alanine; or    b. a mutant JNK3 comprising amino acids 40-402 of SEQ ID NO:3, wherein amino acid 146 is threonine or alanine.    
     
     
         13 . The mutant second kinase according to  claim 12 , wherein in SEQ ID NO:2 amino acid 103 is leucine, amino acid 106 is histidine, amino acid 109 is glycine and amino acid 110 is alanine.  
     
     
         14 . The mutant second kinase according to  claim 12 , wherein in SEQ ID NO:3 amino acid 150 is glycine.  
     
     
         15 . A crystallizable co-complex of a mutant second kinase according to any of claims  10  to 14 and an inhibitor of said first kinase bound to the ATP binding site of said mutant second kinase.  
     
     
         16 . The crystallizable co-complex according to  claim 13 , wherein said first kinase is p38, said second kinase is a MAP kinase and said inhibitor is a pyridinyl-imidazole inhibitor of p38.  
     
     
         17 . The co-complex according to  claim 13 , wherein said pyrindinyl-imidazole inhibitor of p38 is selected from SB203580 or SB202190.