CCI-779 derivatives and methods of making same

Abstract

A method of generating synthetic metabolites of CCI-779 is provided. Five novel CCI-779 derivatives are described, as are methods of using these derivatives for detecting CCI-779 metabolites in samples.

1 . A synthetic CCI-779 reduction and hydroxylation derivative produced by incubating a CYP3A4 isozyme and CCI-779 in the presence of a reducing agent. 
     
     
         2 . An antibody generated using a compound according to  claim 1 . 
     
     
         3 . The antibody according to  claim 2 , wherein the compound is selected from the group consisting of 27-hydroxy seco-CCI-779 (M7), 36-hydroxy CCI-779 (M8), 35-hydroxy CCI-779 (M9), 11-hydroxy CCI-779 (M10), and N-oxide CCI-779 (M12). 
     
     
         4 . A kit for monitoring therapy with CCI-779, said kit comprising a compound according to  claim 1  as a standard. 
     
     
         5 . A kit for monitoring therapy with CCI-779, said kit comprising an antibody according to  claim 5 . 
     
     
         6 . A method for detecting metabolites of CCI-779, said method comprising comparison of a sample to a compound according to  claim 1 . 
     
     
         7 . A method for detecting metabolites of CCI-779, said method comprising detecting binding to an antibody according to  claim 2 . 
     
     
         8 . A method of producing a synthetic metabolite of CCI-779 comprising incubating a CYP3A4 isozyme and CCI-779 in the presence of a reducing agent. 
     
     
         9 . The method according to  claim 8 , wherein the reducing agent is selected from the group consisting of NADH and NADPH. 
     
     
         10 . A method of producing a derivative of CCI-779 comprising incubating a CYP3A4 isozyme and CCI-779 in the presence of a dehydrogenase. 
     
     
         11 . The method according to  claim 10 , wherein the dehydrogenase is glucose-6-phosphate dehydrogenase.