Process for Preparing Imetelstat

Abstract

The present invention relates to a process for preparing the telomerase inhibitor imetelstat using a 3 steps per cycle solid-phase support bound process comprising the steps of deprotection of the 3′-amino group of the support-bound oligonucleotide, coupling with a 5′-phosphoramidite, and sulfurization with an acyl disulfide, characterized by the absence of an additional capping step in each cycle that is used to prevent unreacted 3′-amino oligonucleotide groups from reacting during subsequent cycles. Imetelstat has formula below.

1 . A method of synthesizing the N3′→P5′ thiophosphoramidate oligonucleotide imetelstat of formula 
       
         
           
                 
               
                     
                 
                   imetelstat 
                 
                     
                 
                   
                     
                       
                       
                           
                           
                       
                     
                   
                 
                     
                 
                 
                 
               
                   B 1  = T 
                   B 10  = A 
                 
                   B 2  = A 
                   B 11  = C 
                 
                   B 3  = G 
                   B 12  = A 
                 
                   B 4  = G 
                   B 13  = A 
                 
                   B 5  = G 
                     
                 
                   B 6  = T 
                   T = thymine 
                 
                   B 7  = T 
                   A = adenine 
                 
                   B 8  = A 
                   G = guanine 
                 
                   B 9  = G 
                   C = cytosine 
                 
                     
                 
             
                
                
                
               
               
                
                
               
            
             
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         the method comprises of 
         a) providing a first 3′-amino protected nucleotide attached to a solid-phase support of formula (A) wherein PG is an acid-labile protecting group; 
       
       
         
           
           
               
               
           
         
         b) deprotecting the protected 3′-amino group to form a free 3′-amino group; 
       
       
         
           
           
               
               
           
         
         c) reacting the free 3′-amino group with a protected 3′-aminonucleoside-5′-O-cyanoethyl-N,N-diisopropylaminophosphoramidite monomer of formula (B′ n ) wherein n=2 to form an internucleoside N3′→P5′-phosphoramidite linkage; 
       
       
         
           
           
               
               
           
         
         d) sulfurization of the internucleoside phosphoramidite group using an acyl disulfide to form a N3′→P5′ thiophosphoramidate; 
         e) repeating 11 times in successive order the deprotection step b), the coupling step c) with a protected 3′-aminonucleoside-5′-O-cyanoethyl-N,N-diisopropylamino-phosphoramidite monomer of formula (B′ n ) wherein the protected nucleoside base B′ in monomer (B′ n ) is successively the protected nucleobase B 3  to B 13 in the respective 11 coupling steps, and the sulfurization step d); 
         f) removing the acid-labile protecting group PG; and 
         g) cleaving and deprotecting imetelstat from the solid-phase support; 
         characterized in that no additional capping step is performed in any of the reaction steps a) to e). 
       
     
     
         2 - 13 . (canceled)