Kinase mimic catalysts for asymmetric synthesis of phosphorylated inositols and cycloalkanols

Abstract

The present invention provides peptide-based phosphorylation catalysts (PBPC's) for the asymmetric monophosphorylation of cyclitols, particularly myo-inositols. The PBPC's of the invention effect a regio and enantioselective phosphorylation of a myo-inositol in a manner analogous to enzymatic kinases, thereby functioning as effective “kinase mimics.” Although orders of magnitude less complex in terms of structure than macromolecular proteins, the PBPC's of the invention control product formation with high enantioselectivity (>98% ee). The synthetic (+)-myo-inositol-1-phosphate is optically and spectroscopically equivalent to naturally occurring compound. The ability of the low molecular weight PBPC's of the present invention to mimic stereoselective enzymes represents a powerful approach toward catalytic asymmetric synthesis of biologically important molecules, and for mechanistic modeling of biochemical transformations to enable their use in drug applications.

1 . A compound of the formula  
     
       
         
         
             
             
         
       
     
     wherein R 1  is a lower alkyl; R 2  is an amine protecting group; and Pep is a peptide comprising a peptide residue including at least one natural or non-natural amino acid and having at least one phosphorous moiety.  
   
   
       2 . The compound of  claim 1  wherein R 1 ═C 1  to C 6  straight or branched chain alkyl, R 2  is a carbamate, and Pep is a monopeptide, an oligopeptide or a polypeptide comprising at least one amino acid.  
   
   
       3 . The compound of  claim 2  wherein the carbamate is selected from the group consisting of t-butyl carbamate, 9-fluorenylmethyl carbamate, benzyl carbamate and ortho-nitrobenzyl carbamate.  
   
   
       4 . The compound of  claim 1  wherein R 1  is CH 3  and R 2  is t-butylcarbamate.  
   
   
       5 . The compound of  claim 1  wherein Pep is a an oligopeptide or a polypeptide comprising a sequence of 2 to 50 amino acids.  
   
   
       6 . The compound of  claim 1  wherein Pep is a an oligopeptide or a polypeptide comprising a sequence of 2 to 8 amino acids.  
   
   
       7 . The compound of  claim 1  for catalyzing the stereoselective phosphorylation of cyclitols.  
   
   
       8 . The compound of  claim 7  wherein the cyclitol is an inositol  
   
   
       9 . The compound of  claim 8  wherein the inositol is myo-inositol.  
   
   
       10 . A method of selecting a phosphorylation catalyst of the formula  
     
       
         
         
             
             
         
       
     
     wherein R 1  is a lower alkyl; R 2  is an amine protecting group; and Pep is a peptide comprising a peptide residue including at least one natural or non-natural amino acid for stereoselective phosphorylation of a cyclitol comprising: 
 creating a library having at least one asymmetric phosphorylation catalyst member using an algorithm;  
 chemically synthesizing the phosphorylation catalyst member;  
 subjecting a cyclitol or a partially protected derivative thereof to a phosphorylation reaction with a phosphorylation reagent in the presence of the phosphorylation catalyst member to give a corresponding monophosphate compound; and  
 performing a product assay to determine a stereochemical product composition of the monophosphate compound.  
 
   
   
       11 . The method of  claim 10  wherein R 1 ═C 1  to C 6  straight or branched chain alkyl, R 2  is a carbamate, and Pep is a monopeptide, an oligopeptide or a polypeptide comprising at least one amino acid.  
   
   
       12 . The method of  claim 10  wherein the carbamate is selected from the group consisting of t-butyl carbamate, 9-fluorenylmethyl carbamate, benzyl carbamate and ortho-nitrobenzyl carbamate.  
   
   
       13 . The method of  claim 10  wherein R 1  is CH 3  and R 2  is t-butylcarbamate.  
   
   
       14 . The method of  claim 10  wherein Pep is an oligopeptide or a polypeptide comprising a sequence of 2 to 50 amino acids.  
   
   
       15 . The method of  claim 10  wherein the phosphorylation agent is a dichlorodiakyl phosphate, dichlorodiarylphosphate and derivatives thereof.  
   
   
       16 . The method of  claim 10  wherein the phosphorylation agent is dichlorodiphenyl phosphate.  
   
   
       17 . The method of  claim 10  wherein the cyclitol monophosphate is an inositol monophosphate.  
   
   
       18 . The method of  claim 17  wherein the inositol monophosphate is (D)-myo-inositol-1-phosphate.  
   
   
       19 . The method of  claim 17  wherein the inositol monophosphate is (D)-myo-inositol-3-phosphate.  
   
   
       20 . A method of identifying a phosphorylation catalyst to function as a stereoselective catalyst material for phosphorylation of a cyclitol comprising: 
 chemically synthesizing at least one asymmetric phosphorylation catalyst member;    subjecting a cyclitol or a partially protected derivative thereof to a phosphorylation reaction with a phosphorylation reagent in the presence of the phosphorylation catalyst member to yield a monophosphate compound;    performing a product assay on the monophosphate compound for evaluating a stereochemical product composition of the monophosphate compound;    selecting the phosphorylation catalyst of the formula                          wherein R 1  is a lower alkyl; R 2  is an amine protecting group; and Pep is a peptide.