Specific Targeting of RNA Expanded Repeat Sequences

Abstract

The present invention provides small molecule compounds that can form covalent adducts with specific sequences of RNA, such as the hairpin loop r(CUG) exp sequence which is a cause of myotonic dystrophy type 1 (DM1), or the r(CGG) exp sequence which is a cause of fragile X-associated tremor/ataxia syndrome (FXTAS); to methods of making the small molecule compounds; and to methods of using the small molecular compounds in the treatment of DM1 or of FXTAS in patients afflicted therewith. The invention further provides a method for identifying an RNA target of a small molecule drug in vivo, using a small molecule drug conjugated to an RNA-reactive crosslinker group and a reporter group, contacting a cell or nucleic acid extract with the small molecule drug conjugate, then separating RNA targets crosslinked to the small molecule drug conjugate by interaction of the affinity group with a complementary affinity group.

What is claimed is: 
     
         1 . A compound of formula (I) 
       
         
           
           
               
               
           
         
         wherein
 each individually selected R or R′ is H or (C1-C6)alkyl; 
 n1 is 2, 3, 4, 5, or 6; 
 Z is a (C1-C3)alkylene group, optionally substituted with a reporter or affinity group; 
 L is a linker group comprising an optionally substituted (C1-C6)alkylene, wherein one or two carbon atoms is optionally replaced by O; 
 XL is an RNA-reactive crosslinking group; 
 each individually selected Ht is a group of formula 
 
       
       
         
           
           
               
               
           
         
         wherein
 n2 is 0, 1, 2, or 3; 
 n3 is 0, 1, 2, or 3; 
 Y is O or CH 2 ; 
 a wavy line indicates a point of bonding; 
 or any tautomer thereof; or any salt thereof, or any stereoisomer thereof. 
 
       
     
     
         2 . The compound of  claim 1  wherein the crosslinking group is an alkylating group. 
     
     
         3 . The compound of  claim 2  wherein the alkylating group is a chlorambucil derivative. 
     
     
         4 . The compound of  claim 1  wherein the crosslinking group is a triggered crosslinker using photochemical or shape-triggered catalysis. 
     
     
         5 . The compound of  claim 1  wherein the compound is of formula 
       
         
           
           
               
               
           
         
         or is of formula 
       
       
         
           
           
               
               
           
         
         wherein Ht, R, and R′, are as defined in  claim 1 ;
 or any tautomer thereof; or any salt thereof, or any stereoisomer thereof. 
 
       
     
     
         6 . The compound of  claim 1  wherein Ht is 
       
         
           
           
               
               
           
         
         wherein a wavy line indicates a point of bonding; 
         R is n-propyl; R′ is H; or any combination thereof; 
         or any tautomer thereof; or any salt thereof, or any stereoisomer thereof. 
       
     
     
         7 . The compound of  claim 1  wherein the compound is 2H-4-CA of formula 
       
         
           
           
               
               
           
         
         or is compound 2H-5-CA of formula 
       
       
         
           
           
               
               
           
         
         wherein Ht is a group of formula 
       
       
         
           
           
               
               
           
         
         wherein a wavy line indicates a point of bonding; 
         or any tautomer thereof; or any salt thereof, or any stereoisomer thereof. 
       
     
     
         8 . The compound of  claim 1  wherein Z is an unsubstituted alkylene group. 
     
     
         9 . The compound of  claim 1  wherein Z is an alkylene group substituted with an affinity group. 
     
     
         10 . The compound of  claim 9  wherein the affinity group is a biotin-comprising moiety. 
     
     
         11 . The compound of  claim 10 , wherein the compound is 
       
         
           
           
               
               
           
         
         wherein Ht, R, R′, n1, L, and XL, are as defined in  claim 1 ; 
         or any tautomer thereof; or any salt thereof, or any stereoisomer thereof. 
       
     
     
         12 . The compound of  claim 11 , wherein n1 is 4 or 5. 
     
     
         13 . The compound of  claim 12 , wherein the compound is 2H-4-CA-biotin of formula 
       
         
           
           
               
               
           
         
         or is 2H-5-CA-biotin of formula 
       
       
         
           
           
               
               
           
         
         or any tautomer thereof; or any salt thereof, or any stereoisomer thereof. 
       
     
     
         14 . A covalently-linked complex of the compound of formula (I) of  claim 1  and a segment of RNA comprising a hairpin loop. 
     
     
         15 . The complex of  claim 14  wherein the RNA comprises an r(CUG) exp  segment. 
     
     
         16 . The complex of  claim 14  wherein the RNA comprises an r(CGG) exp  segment.