RNA Modulating Oligonucleotides with Improved Characteristics for the Treatment of Duchenne and Becker Muscular Dystrophy

Abstract

The current invention provides an improved oligonucleotide and its use for treating, ameliorating, preventing and/or delaying DMD or BMD.

1 . An oligonucleotide comprising a 2′-O-methyl RNA monomer and a phosphorothioate backbone and comprising a 5-methyluracil and/or a 5-methylcytosine and/or a 2,6-diaminopurine base, said oligonucleotide being able to induce skipping of an exon of the dystrophin pre-mRNA. 
     
     
         2 . An oligonucleotide according to  claim 1 , wherein said oligonucleotide comprises a 5-methylcytosine and/or a 5-methyluracil base. 
     
     
         3 . An oligonucleotide according to  claim 1 , wherein said oligonucleotide comprises a 2,6-diaminopurine base. 
     
     
         4 . An oligonucleotide according to  claim 1 , wherein said oligonucleotide has an improved parameter by comparison to a corresponding oligonucleotide comprising a 2′-O-methyl RNA monomer and a phosphorothioate backbone without a 5-methylcytosine, and a 5-methyluracil and a 2,6-diaminopurine wherein said improved parameter is selected from the group consisting of: increased binding affinity, longer half-life, increased exon skipping activity, increased biostability, wider (intratissue) distribution, increased cellular uptake, increased trafficking, and/or lower immunogenicity. 
     
     
         5 . An oligonucleotide according to  claim 1 , wherein the length of said oligonucleotide is less than 34 nucleotides. 
     
     
         6 . An oligonucleotide according to  claim 1 , wherein said oligonucleotide is reverse complementary to and/or binds to and/or targets and/or hybridizes with at least a part of a dystrophin exon and/or non-exon region. 
     
     
         7 . An oligonucleotide according to  claim 1 , wherein said oligonucleotide comprises or consists of a sequence which is reverse complementary to and/or binds to and/or targets and/or hybridizes at least a part of dystrophin pre-mRNA exons 44 to 55, said oligonucleotide part having from 10 to 33 nucleotides. 
     
     
         8 . An oligonucleotide according to  claim 7 , wherein said oligonucleotide comprises a 2′-O-methyl RNA monomer and a phosphorothioate backbone, said oligonucleotide is represented by a nucleotide or a base sequence comprising or consisting of one of SEQ ID NO: 52, 14-51, 53-90 or by a nucleotide sequence comprising or consisting of a fragment of one of SEQ ID NO: 52, 14-51, 53-90 and said oligonucleotide comprises a 5-methyluracil and/or a 5-methylcytosine and/or a 2,6-diaminopurine base. 
     
     
         9 . An oligonucleotide according to  claim 8 , wherein said oligonucleotide is represented by a nucleotide or a base sequence comprising or consisting of one of SEQ ID NO: 52, 15, 21, 31, 40, 57, or by a nucleotide or a base sequence comprising or consisting of a fragment of one of SEQ ID NO: 52, 15, 21, 31, 40, 57. 
     
     
         10 . An oligonucleotide according to  claim 8 , wherein said oligonucleotide is represented by a nucleotide or a base sequence comprising or consisting of one of SEQ ID NO:92, 171-215, 217, 218, 219 or by a nucleotide or a base sequence comprising or consisting of a fragment of one of SEQ ID NO: 92, 171-215, 217, 218, 219. 
     
     
         11 . An oligonucleotide according to  claim 10 , wherein said oligonucleotide is represented by a nucleotide or a base sequence comprising or consisting of one of SEQ ID NO:92, 171, 173, 185, 187, 200, 206, 207, 208, 210, 213, 217, 218 or 219 or by a nucleotide or a base sequence comprising or consisting of a fragment of one of SEQ ID NO: 92, 171, 173, 185, 187, 200, 206, 207, 208, 210, 213, 217, 218 or 219 said fragment comprising or consisting of at least 10 contiguous nucleotides or bases of SEQ ID NO: 92, 171, 173, 185, 187, 200, 206, 207, 208, 210, 213, 217, 218 or 219 and such fragment having a length of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 or 33 nucleotides. 
     
     
         12 . A composition comprising an oligonucleotide as defined in  claim 1 . 
     
     
         13 . A composition according to  claim 12 , comprising at least one excipient that may enhance the targeting and/or delivery of said composition and/or said oligonucleotide to a tissue and/or cell and/or into a tissue and/or cell. 
     
     
         14 . A method for preventing, treating, and/or delaying Duchenne Muscular Dystrophy or Becker Muscular Dystrophy by administering an oligonucleotide as defined in  claim 1  to a subject in the need thereof. 
     
     
         15 . A method for preventing, treating, and/or delaying Duchenne Muscular Dystrophy or Becker Muscular Dystrophy by administering a composition as defined in  claim 12  to a subject in the need thereof.