US2004097722A9PendingUtilityA9

Preparation of phosphorothioate oligomers

Assignee: UNIV MCGILLPriority: Oct 20, 1995Filed: Apr 25, 2003Published: May 20, 2004
Est. expiryOct 20, 2015(expired)· nominal 20-yr term from priority
C07F 9/65846C07H 9/04C07H 9/06C07F 9/1411C07H 19/10C07H 21/00
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Claims

Abstract

Methods and intermediates for the preparation of oligomers containing diastereomerically enriched phosphorothioate linkages are disclosed.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . A method for synthesizing an oligomer having a phosphorothioate linkage comprising the steps of:  
       reacting a first synthon of Formula I:  
       
         
           
           
               
               
           
         
       
       wherein: 
 Q is independently O or S;  
 R 1  is a hydroxyl protecting group;  
 R 2  is a chiral auxiliary of formula —C(R 8 )R 3 —C(R 16 )R 5 —CHR 6 —NHR 7 ;  
 R 3  is hydrogen, alkyl, cyanomethyl, monohalomethyl, dihalomethyl, trigalomethyl, —CH 2 Si(R 4 ) 3 , or —CH 2 —SO k R 4  where k is 0, 1 or 2;  
 R 4  is independently alkyl, aryl, aralkyl or alkaryl having up to 15 carbon atoms;  
 R 5  is H, —CN, —Si(R 4 ) 3 , SO k R 4  or halogen;  
 or R 8  and R 16  are each H, and R 3  and R 5 , together, form one of the structures  
                     
  wherein: 
 R 10  and R 11  are H, alkyl having from 1 to about 10 carbons, —CH 2 C(═O)OR 22 , —CH 2 CN, —CH 2 Si(CH 3 ) 3 , or o- or p-C 6 H 4 —R 21 ; 
 R 21  is hydrogen, —O—C(═O)CH 3 , alkoxy having from 1 to about 10 carbons, —NO 2 , or —N(R 22 ) 2 ;  
 R 22  is independently H or alkyl having from one to about 10 carbon atoms;  
 p is 1 or 2;  
 
 Z 1  and Z 2  are independently halogen, CN, —Si(CH 3 ) 3 , and —C(═O)OR 22 ;  
 R 30  is hydrogen, —O—C(═O)CH 3 , alkoxy having from 1 to about 10 carbons, or —O—Si(R 4 ) 3 ;  
 
 R 6  is H, alkyl or aralkyl having up to 15 carbon atoms;  
 or R 5  and R 6 , together with the atoms to which they are attached, form a 5 or 6 membered ring;  
 R 7  is alkyl or aralkyl having up to 15 carbon atoms;  
 or R 6  and R 7 , together, form one of the structures  
                     wherein V, T, and Z are independently CH or N;    R 8  is H or methyl;    R 16  is H, alkyl or aralkyl having up to 15 carbon atoms;    B a is a nucleobase; and    n is an integer from 0 to 50;    
 with a second synthon of Formula II:  
                     
  wherein: 
 R 9  is a hydroxyl protecting group or a linker connected to a solid support; and  
 m is an integer from 0 to 50; for a time and under reaction conditions effective to form a third synthon of Formula III:  
                     
 and  
 contacting said third synthon with a sulfurizing agent to form an oligomer of Formula IV:  
                     
 wherein D is said phosphorothioate linkage having the formula:  
                     
 
 
     
     
         2 . The method of  claim 1  wherein said phosphorothioate linkage is diastereomerically enriched.  
     
     
         3 . The method of  claim 1  wherein about 75% of said phosphorothioate linkage is in a single stereoisomeric form.  
     
     
         4 . The method of  claim 1  wherein about 85% of said phosphorothioate linkage is in a single stereoisomeric form.  
     
     
         5 . The method of  claim 1  wherein about 95% of said phosphorothioate linkage is in a single stereoisomeric form.  
     
     
         6 . The method of  claim 1  wherein said phosphorothioate linkage is in a single stereoisomeric form, substantially free of other stereoisomeric forms.  
     
     
         7 . The method of  claim 1  wherein n is 0.  
     
     
         8 . The method of  claim 1  wherein said first synhon is in a single stereoisomeric form, substantially free of other stereoisomeric forms.  
     
     
         9 . The method of  claim 1  further comprising removing said R 1  group from said phosphorothioate.  
     
     
         10 . The method of  claim 1  further comprising removing said chiral auxiliaries.  
     
     
         11 . The method of  claim 1  wherein said compound of Formula IV contains a plurality of said phosphorothioate linkages.  
     
     
         12 . The method of  claim 1  wherein said first and second synthons are reacted at a temperature of from about −20° C. to about 40° C.  
     
     
         13 . The method of  claim 1  wherein said first and second synthons are reacted at a temperature of from about −15° C. to about 0° C.  
     
     
         14 . The method of  claim 1  wherein said first synthon is formed by reacting a compound of Formula V:  
       
         
           
           
               
               
           
         
       
       with an azaphospholane of Formula VIa:  
       
         
           
           
               
               
           
         
       
       wherein R 3 -R 8  are as defined above; and X is halogen, dialkylamino, inidazols, triazole or substituted phenoxy wherein said substituents are electron withdrawing.  
     
     
         15 . The method of  claim 14  wherein said electron withdrawing substituents are halogen or nitro.  
     
     
         16 . The method of  claim 14  wherein said azaphospholane is produced by reacting a reagent of formula HO—C(R 8 )R 3 —C(R 16 )R 5 —CHR 6 —NHR 7  and a phosphorus trihalide, phosphorus tri(dialkylamide), phosphorus triphenoxide, or phosphorus tri(imidazolides).  
     
     
         17 . The method of  claim 1  wherein said first synthon is formed by reacting a compound of Formula VII:  
       
         
           
           
               
               
           
         
       
       and a γ-amino alcohol of formula HO—C(R 8 )R 3 —C(R 16 )R 5 —CHR 6 —NHR 7 ; wherein X and R 1 -R 8  and R 16  are as defined above.  
     
     
         18 . The method of  claim 17  wherein X is chlorine.  
     
     
         19 . The method of  claim 17  wherein said reaction is stereoselective.  
     
     
         20 . The method of  claim 18  wherein said first synthon is in a single stereoisomeric form, substantially free of other stereoisomeric forms.  
     
     
         21 . The method of  claim 1  wherein'said reaction is performed in the presence of a catalyst.  
     
     
         22 . The method of  claim 21  wherein said catalyst has one of the Formulas VIII or IX:  
       
         
           
           
               
               
           
         
       
       wherein: 
 R 12  and R 13  are independently hydrogen, halogen, cyano, nitro, alkyl having from one to 10 carbons, substituted alkyl having from one to 10 carbons, and ester group, or R 12  and R 13  together with the carbon atoms to which they are attached, form a substituted or unsubstituted phenyl ring where said substituents are electron withdrawing; and  
 R 14  is hydrogen, halogen, cyano, nitro, thio, alkyl having from one to 10 carbons, substituted alkyl having from one to 10 carbons, norbornyl, substituted norbornyl, aryl, substituted aryl wherein said substituents are electron withdrawing, or has the formula:  
                     
 wherein L is protecting group.  
 
     
     
         23 . The method of  claim 21  wherein R 14  is halogen or nitro, and wherein R 12  and R 13  are each halogen or each cyano.  
     
     
         24 . The method of  claim 22  wherein R 14  is bromine, and R 12  and R 13  are each cyano.  
     
     
         25 . The method of  claim 22  wherein R 14 , R 12  and R 13  are each bromo.  
     
     
         26 . The method of  claim 21  wherein R 14  has one of the formulas:  
       
         
           
           
               
               
           
         
       
       wherein R 15  is H, methyl, trialkylsilyl or acetyl.  
     
     
         27 . The method of  claim 1  wherein R 3  is cyanomethyl or —CH 2 —SO k R 4  where k is 0, 1 or 2.  
     
     
         28 . The method of  claim 27  wherein R 7  is lower alkyl or aralkyl.  
     
     
         29 . The method of  claim 1  wherein said first synthon has one of the Formulas Xa, XIa, XIIa, XIIIa or XXa:  
       
         
           
           
               
               
           
         
       
       wherein W has the formula:  
       
         
           
           
               
               
           
         
       
       and R 1 -R 16 , V, T and Z are as defined above.  
     
     
         30 . The method of  claim 29  wherein said first synthon has the Formula Xb or Xc:  
       
         
           
           
               
               
           
         
       
     
     
         31 . The method of  claim 29  where;n said first synthon has the Formula XIVa:  
       
         
           
           
               
               
           
         
       
       wherein p, Z 1  and Z 2  are as defined above.  
     
     
         32 . The method of  claim 31  wherein said first synthon has the Formula XVa or XVIa:  
       
         
           
           
               
               
           
         
         wherein Y, R 7 , Z 1 , Z 2 , and p are as defined above.  
       
     
     
         33 . The method of  claim 29  wherein said first synthon has the Formula XVIIa or XVIIIa:  
       
         
           
           
               
               
           
         
       
       wherein W, R 7 , R 10  and R 11  are as defined above.  
     
     
         34 . The method of  claim 1  further comprising removing said R 9  groups.  
     
     
         35 . The method of  claim 1  further comprising removing said R 2  groups.  
     
     
         36 . The oligomer produced by the method of  claim 1 .  
     
     
         37 . An azaphospholane of Formula VIb:  
       
         
           
           
               
               
           
         
       
       wherein: 
 y is X or W wherein X is halogen, dialkylamino, imidazole, triazole or substituted phenoxy wherein said substituents are electron withdrawing, and W has the formula:  
                     
 wherein:  
 Q is independently O or S;  
 R 1  is a hydroxyl Protecting group;  
 R 2  is a chiral auxiliary of formula —C(R 8 )R 3 —C(R 16 )R 5 —CHR 6 —NHR 7 ;  
 R 3  is hydrogen, alkyl, cyanomethyl, monohalomethyl, dihalomethyl, trihalomethyl, —CH 2 Si(R 4 ) 3 , or —CH 2 —SO k R 4  where k is 0, 1 or 2;  
 R 4  is independently alkyl, aryl, aralkyl or alkaryl having up to 15 carbon atoms;  
 R 5  is H, —CN, —Si(R 4 ) 3 , SO k R 4  or halogen;  
 or R 8  and R 16  are each H, and R 3  and R 5 , together, form one of the structures  
                     
  wherein: 
 R 10  and R 11  are H, alkyl having from 1 to about 10 carbons, —CH 2 C(═O)OR 22 , —CH 2 CN, —CH 2 Si(CH 3 ) 3 , or o- or p-C 6 H 4 —R 21 ;  
 R 21  is hydrogen, —O—C(═O)CH 3 , alkoxy having from 1 to about 10 carbons, —NO 2 , or —N(R 22 ) 2 ;  
 R 22  is independently H or alkyl having from one to about 10 carbon atoms;  
 p is 1 or 2;  
 Z 1  and Z 2  are independently halogen, CN, —Si(CH 3 ) 3 , and —C(═O)OR 22 ;  
 R 30  is hydrogen, —O—C(═O)CH 3 , alkoxy having from 1 to about 10 carbons, or —O—Si(R 4 ) 3 ;  
 
 R 6  is H, alkyl or aralkyl having up to 15 carbon atoms;  
 or R 5  and R 6 , together with the atoms to which they are attached, form a 5 or 6 membered ring;  
 R 7  is alkyl or aralkyl having up to 15 carbon atoms;  
 or R 6  and R 7 , together, form one of the structures  
                     
 wherein X, T, and Z are independently CH or NH; or Z and R 5 , together with the atoms to which they are attached, form a phenyl ring;  
 R 8  is H or methyl;  
 R 9  is a hydroxyl protecting group or a solid support;  
 R 16  is H, alkyl or aralkyl having up to 15 carbon atoms;  
 B is a nucleobase; and  
 n is an integer from 0 to 50.  
 
     
     
         38 . The azaphospholane of  claim 37  wherein Y is W and n is 0.  
     
     
         39 . The azaphospholane of  claim 37  wherein Y is halogen.  
     
     
         40 . The azaphospholane of  claim 37  wherein Y is chlorine.  
     
     
         41 . The azaphospholane of  claim 37  wherein R 7  is alkyl.  
     
     
         42 . The azaphospholane of  claim 37  wherein R 7  R 6  and the atoms to which they are attached form a 5 or 6 membered ring.  
     
     
         43 . The azaphospholane of  claim 38  wherein R 3  is cyanomethyl or —CH 2 —SO k R 6  where k is 0, 1 or 2.  
     
     
         44 . The azaphospholane of  claim 37  having one of the Formulas XVIc or XVId:  
       
         
           
           
               
               
           
         
       
     
     
         45 . The azaphospholane of  claim 36  having one of the Formulas Xb, XIb, XIIb, XIIIb or XXb:  
       
         
           
           
               
               
           
         
       
       wherein R 3 -R 16 , Y, V, T, Z, Z 1 , Z 2  and p are as defined above.  
     
     
         46 . The azaphosphalane of  claim 45  having the Formula XIVb:  
       
         
           
           
               
               
           
         
       
       wherein Y, R 6 , R 7 , R 8 , p, Z 1  and Z 2  are as defined above.  
     
     
         47 . The azaphosphalane of  claim 44  having the Formula Xd or Xe:  
       
         
           
           
               
               
           
         
       
       wherein Y, R 3 , R 5 , R 8 , and R 1 6 are as defined above.  
     
     
         48 . The azaphospholane of  claim 45  having the Formula XVIIb or XVIIIb.  
       
         
           
           
               
               
           
         
       
       wherein Y, R 7 , R 10  and R 11  are as defined above.  
     
     
         49 . The azaphospholane of  claim 45  having the Formula XVb or XVIb:  
       
         
           
           
               
               
           
         
       
       wherein Y, R 7 , Z 1 , Z 2 , and p are as defined above.  
     
     
         50 . The azaphospholane of  claim 37  wherein said azaphospholane is diastereomerically enriched.  
     
     
         51 . The azaphospholane of  claim 37  wherein 75% of said azaphospholane is in a single stereoisomeric form.  
     
     
         52 . The azaphospholane of  claim 37  wherein 85% of said azaphospholane is in a single stereoisomeric form.  
     
     
         53 . The azaphospholane of  claim 37  wherein 95% of said azaphospholane is in a single stereoisomeric form.  
     
     
         54 . The azaphospholane of  claim 37  wherein said azaphospholane is in a single stereoisomeric form, substantially free of other stereoisomeric forms.  
     
     
         55 . An oligomeric compound comprising a moiety having a phosphite linkage of the Formula XXX:  
       
         
           
           
               
               
           
         
       
       wherein: 
 R 3  is hydrogen, alkyl, cyanomethyl, monohalomethyl, dihalomethyl, trihalomethyl, —CH 2 Si(R 4 ) 3 , or —CH 2 —SO k R 4  where k is 0, 1 or 2;  
 R 4  is independently alkyl, aryl, aralkyl or alkaryl having up to 15 carbon atoms;  
 R 5  is H, —CN, —Si(R 4 ) 3 , SO k R 4  or halogen;  
 or R 8  and R 16  are each H, and R 3  and R 3 , together, form one of the structures  
                     
  wherein: 
 R 10  and R 11  are H, alkyl having from 1 to about 10 carbons, —CH 2 C(═O)OR 22 , —CH 2 CN, —CH 2 Si(CH 3 ) 3 , or o- or p-C 6 H 4 —R 21 ;  
 R 21  is hydrogen, —O—C(═O)CH 3 , alkoxy having from 1 to about 10 carbons, —NO 2 , or —N(R 22 ) 2 ;  
 R 22  is independently H or alkyl having from one to about 10 carbon atoms;  
 p is 1 or 2;  
 Z 1  and Z 2  are independently halogen, CN, —Si(CH 3 ) 3 , and —C(═O)OR 22 ;  
 R 30  is hydrogen, —O—C(═O)CH 3 , alkoxy having from 1 to about 10 carbons, or —O—Si(R 4 ) 3 ;  
 
 R 6  is H, alkyl or aralkyl having up to 15 carbon atoms;  
 or R 5  and R 6 , together with the atoms to which they are attached, form a 5 or 6 membered ring;  
 R′ is alkyl or aralkyl having up to 15 carbon atoms;  
 or R 6  and R 7 , together, form one of the structures  
                     
 wherein X, T, and Z are independently CH or NH; or Z and R 5 , together with the atoms to which they are attached, form a phenyl ring;  
 R 8  is H or methyl;  
 R 9  is a hydroxyl protecting group or a solid support;  
 R 16  is H, alkyl or aralkyl having up to 15 carbon atoms;  
 B is a nucleobase; and  
 n is an integer from 0 to 50.  
 
     
     
         56 . The oligomeric compound of  claim 55  wherein 75% of said phosphosphite linkage is in a single stereoisomeric form.  
     
     
         57 . The oligomeric compound of  claim 55  wherein 85% of said phosphosphite linkage is in a single stereoisomeric form.  
     
     
         58 . The oligomeric compound of  claim 55  wherein 95% of said phosphosphite linkage is in a single stereoisomeric form.  
     
     
         59 . The oligomeric compound of  claim 55  wherein said phosphosphite linkage is in a single stereoisomeric form, substantially free of other stereoisomeric forms.

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