Antisense oligonucleotides for inducing exon skipping and methods of use thereof

Abstract

An antisense molecule capable of binding to a selected target site to induce exon skipping in the dystrophin gene, as set forth in SEQ ID NO: 1 to 202.

1 . An antisense molecule capable of binding to a selected target site to induce exon skipping in the dystrophin gene, as set forth in SEQ ID NO: 1 to 202. 
     
     
         2 . An antisense molecule according to  claim 1  capable of inducing exon skipping in exons 3, 4, 8, 10 to 16, 19 to 40, 42 to 44, 46, 47 and 50 to 53 of the dystrophin gene. 
     
     
         3 . A combination of two or more antisense molecules according to  claim 1  or  2  capable of binding to a selected target to induce exon skipping in the dystrophin gene. 
     
     
         4 . A combination or two or more antisense molecules according to  claim 3  selected from Table 1B. 
     
     
         5 . A combination of two or more antisense molecules according to  claim 1  or  2  joined together to form a “weasel”, wherein said weasel is capable of binding to a selected target to induce exon skipping in the dystrophin gene. 
     
     
         6 . A combination of two or more antisense molecules according to  claim 5  selected from Table 1C. 
     
     
         7 . The antisense molecule according to any one of  claims 1  to  6 , capable of binding to a selected target site, wherein the target site is an rnRNA splicing site selected from a splicer donor site, splice acceptor sites or exonic splicing enhancer elements. 
     
     
         8 . A method of treating muscular dystrophy in a patient comprising administering to the patient a composition comprising an antisense molecule according to anyone of  claims 1  to  6 . 
     
     
         9 . A pharmaceutical or therapeutic composition for the treatment of muscular dystrophy in a patient comprising (a) at least an antisense molecule according to any one of  claims 1  to  6 , and (b) one or more pharmaceutically acceptable carriers and/or diluents. 
     
     
         10 . The composition according to  claim 9 , comprising about 20 nM to 600 nM of the antisense molecule. 
     
     
         11 . The use of an antisense molecule according to any one of  claims 1  to  6  for the manufacture of a medicament for modulation of muscular dystrophy. 
     
     
         12 . An antisense molecule according to any one of  claims 1  to  6  for use in antisense molecule based therapy. 
     
     
         13 . An antisense molecule according to any one of  claims 1  to  6  as herein before described with reference to the examples. 
     
     
         14 . A kit comprising at least one antisense molecule according to any one of  claims 1  to  6 , a suitable carrier and instructions for its use. 
     
     
         15 . An oligomer for ameliorating DMD, the oligomer comprising at least 25 contiguous bases of a base sequence selected from the group consisting of: 
       
         
           
                 
                 
               
                     
                   a) CXG XXG CCX CCG GXX CXG AAG GXG; 
                 
                     
                     
                 
                     
                   b) C AXX CAA CXG XXG CCX CCG GXX CXG AAG GXG; 
                 
                     
                   and 
                 
                     
                     
                 
                     
                   c) XXG CCX CCG GXX CXG AAG GXG, 
                 
             
                
                
                
                
                
                
               
            
           
         
       
       wherein X=U or T, wherein the oligomer's base sequence can vary from the above sequence at up to two base positions, and wherein the molecule can bind to a target site to cause exon skipping in an exon of the dystrophin gene. 
     
     
         16 . The oligomer according to  claim 15 , wherein the exon of the dystrophin gene at which exon skipping is caused is exon 53. 
     
     
         17 . The oligomer according to  claim 15 , wherein the oligomer causes an exon skipping rate of at least 50%. 
     
     
         18 . The oligomer according to  claim 15 , wherein the oligomer is between 25 and 35 bases in length. 
     
     
         19 . The oligomer according to  claim 15 , wherein the oligomer is 30 bases in length. 
     
     
         20 . The oligomer according to  claim 15 , wherein the oligomer is conjugated to or complexed with a distinct chemical entity. 
     
     
         21 . The oligomer according to  claim 15 , wherein the oligomer is a phosphorodiamidate morpholino oligonucleotide (PMO). 
     
     
         22 . A vector for ameliorating DMD, the vector encoding an oligomer according to  claim 15 , wherein when introduced into a human cell the oligomer is expressed. 
     
     
         23 . A pharmaceutical composition for ameliorating DMD, the composition comprising an oligomer according to  claim 15  or a vector according to  claim 22 , and a pharmaceutically acceptable carrier, adjuvant or vehicle. 
     
     
         24 . A pharmaceutical composition according to  claim 23  comprising a plurality of oligomers or vectors encoding oligomers, or a combination of the oligomers and vectors, wherein the oligomers and/or vectors in the pharmaceutical composition cause skipping in a plurality of exons.