Synthesis of Cyclosporin Analogs

Abstract

The invention is directed to isomeric mixtures of cyclosporine analogues that are structurally similar to cyclosporine A. The mixtures possess enhanced efficacy and reduced toxicity over the individual isomers and over naturally occurring and other presently known cyclosporines and cyclosporine derivatives. Embodiments of the present invention are directed toward cis and trans-isomers of cyclosporin A analogs referred to as ISA TX 247, and derivatives thereof. ISA TX 247 isomers and alkylated, arylated, and deuterated derivatives are synthesized by stereoselective pathways where the particular conditions of a reaction determine the degree of stereoselectivity. Stereoselective pathways may utilize a Wittig reaction, or an organometallic reagent comprising inorganic elements such as boron, silicon, titanium, and lithium. The ratio of isomers in a mixture may range from about 10 to 90 percent by weight of the (E)-isomer to about 90 to 10 percent by weight of the (Z)-isomer, based on the total weight of the mixture.

1 . A method of preparing an isomeric mixture of cyclosporin A analogs modified at the 1-amino acid residue, wherein the synthetic pathway comprises the steps of:
 a) protecting the β-alcohol of cyclosporin A by forming trimethylsilyl cyclosporine;   b) oxidizing the trimethylsilyl cyclosporin A with ozone as the oxidizing agent following by work-up with a reducing agent;   c) converting an intermediate trimethylsilyl cyclosporin A aldehyde to a mixture of (E) and (Z)-isomers of trimethylsilyl cyclosporin A-1,3-diene by reacting the intermediate with a phosphorus glide prepared from a tributylallylphosphonium halide or triphenylphosphonium halide via a Witting reaction, optionally in the presence of a lithium halide; and   d) preparing a mixture of (E) and (Z)-isomers of cyclosporin A analogs modified at the 1-amino acid residue by deprotecting the mixture of (E) and (Z)-isomers of trimethylsilyl cyclosporin A-1,3-diene.   
     
     
         2 . The method of  claim 1 , wherein the halide is bromide. 
     
     
         3 . The method of  claim 1 , wherein step c) is carried out in a solvent that comprises tetrahydrofuran and/or toluene used in the presence of a sodium or potassium lower alkoxide, or a carbonate, at a temperature between about −80° C. and about 110° C. 
     
     
         4 . The method of  claim 1 , wherein the sodium or potassium lower alkoxide is potassium-tert-butoxide. 
     
     
         5 . The method of  claim 1 , wherein step c) is carried out with a reagent selected from the group consisting of hydrochloric acid, acetic acid, citric acid, a Lewis acid, and HF-based reagents. 
     
     
         6 . The method of  claim 1 , wherein the oxidizing step is carried out with ozone and is an ozonolysis and further wherein the oxonolysis also comprises treatment with a reducing agent. 
     
     
         7 . The method of  claim 6 , wherein the ozonolysis is done at a temperature ranging from about −80° C. to about 0° C. 
     
     
         8 . The method of  claim 6 , wherein the reducing agent is selected from the group consisting of trialkyl phosphines, triaryl phosphines, and trialkylamines. 
     
     
         9 . The method of  claim 6 , wherein the reducing agent is selected from the group consisting of alkylaryl sulfides, thiosulfates, and dialkyl sulfides. 
     
     
         10 . The method of  claim 9 , wherein the reducing agent is dimethyl sulfide. 
     
     
         11 . The method of  claim 8 , wherein the reducing agent is tributyl phosphine. 
     
     
         12 . The method of  claim 8 , wherein the reducing agent is a trialkylamine. 
     
     
         13 . The method of  claim 12 , wherein the reducing agent is triethylamine. 
     
     
         14 . The method of  claim 6 , wherein a solvent used for the ozonolysis is a lower alcohol. 
     
     
         15 . The method of  claim 14 , wherein the lower alcohol solvent is methanol.