Antisense Oligonucleotides for Treatment of Spinal Muscular Atrophy

Abstract

Various aspects of the present invention are directed to compounds targeted to various regions of the survival motor neuron 2 (SMN2) gene. Such compounds may be used to increase incorporation of exon 7 in processed transcripts of SMN2. Such compounds may therefore be useful in increasing the amount of full-length SMN protein produced by the SMN2 gene. As such, these compounds may provide a therapeutic approach for treatment of spinal muscular atrophy (SMA).

What is claimed is: 
     
         1 . An antisense oligonucleotide having a nucleotide sequence that is complementary to a portion of a sequence of intron 6, intron 7, or exon 7 of a survival motor neuron (SMN) 2 gene, the nucleotide sequence of the antisense oligonucleotide being chosen from [SEQ ID NO 1], [SEQ ID NO 2], [SEQ ID NO 4], [SEQ ID NO 5], [SEQ ID NO 6], [SEQ ID NO 7], [SEQ ID NO 8], [SEQ ID NO 9], [SEQ ID NO 10], [SEQ ID NO 11], [SEQ ID NO 12], [SEQ ID NO 13], [SEQ ID NO 14], [SEQ ID NO 15], [SEQ ID NO 16], and [SEQ ID NO 17]. 
     
     
         2 . The antisense oligonucleotide of  claim 1 , wherein each nucleoside of the antisense oligonucleotide is linked to a morpholino ring. 
     
     
         3 . The antisense oligonucleotide of  claim 2 , wherein morpholino rings are linked to one another via phosphorodiamidate groups. 
     
     
         4 . The antisense oligonucleotide of  claim 1 , wherein the nucleotide sequence is targeted to a cis splicing regulatory element. 
     
     
         5 . The antisense oligonucleotide of  claim 4 , wherein the cis splicing regulatory element is chosen from an exonic splicing enhancer, an exonic splicing silencer, an intronic splicing enhancer, and an intronic splicing silencer. 
     
     
         6 . The antisense oligonucleotide of  claim 4 , wherein the cis splicing regulatory element is an intronic splicing silencer. 
     
     
         7 . The antisense oligonucleotide of  claim 1 , wherein the nucleotide sequence of the antisense oligonucleotide is complementary to a continuous portion of a sequence of intron 6, intron 7, or exon 7 of a survival motor neuron (SMN) 2 gene. 
     
     
         8 . The antisense oligonucleotide of  claim 1 , wherein the nucleotide sequence of the antisense oligonucleotide is complementary to a non-continuous portion of a sequence of intron 6, intron 7, or exon 7 of a survival motor neuron (SMN) 2 gene. 
     
     
         9 . A method of incorporating exon 7 of the SMN2 gene into transcripts of the SMN2 gene, the method comprising:
 administering to a subject an antisense oligonucleotide having a nucleotide sequence:
 (i) that is chosen from [SEQ ID NO 1], [SEQ ID NO 2], [SEQ ID NO 4], [SEQ ID NO 5], [SEQ ID NO 6], [SEQ ID NO 7], [SEQ ID NO 8], [SEQ ID NO 9], [SEQ ID NO 10], [SEQ ID NO 11], [SEQ ID NO 12], [SEQ ID NO 13], [SEQ ID NO 14], [SEQ ID NO 15], [SEQ ID NO 16], and [SEQ ID NO 17]; or 
 (ii) which is sufficiently homologous to [SEQ ID NO 1], [SEQ ID NO 2], [SEQ ID NO 4], [SEQ ID NO 5], [SEQ ID NO 6], [SEQ ID NO 7], [SEQ ID NO 8], [SEQ ID NO 9], [SEQ ID NO 10], [SEQ ID NO 11], [SEQ ID NO 12], [SEQ ID NO 13], [SEQ ID NO 14], [SEQ ID NO 15], [SEQ ID NO 16], or [SEQ ID NO 17]; 
   such that the nucleotide sequence of the antisense oligonucleotide is complementary to a portion of a sequence of intron 6, intron 7, or exon 7 of the SMN2 gene.   
     
     
         10 . The method of  claim 9 , wherein each nucleoside of the antisense oligonucleotide are linked to a morpholino ring. 
     
     
         11 . The method of  claim 10 , wherein morpholino rings are linked to one another via phosphorodiamidate groups. 
     
     
         12 . The method of  claim 9 , wherein the nucleotide sequence is targeted to a cis splicing regulatory element. 
     
     
         13 . The method of  claim 12 , wherein the cis splicing regulatory element is chosen from an exonic splicing enhancer, an exonic splicing silencer, an intronic splicing enhancer, and an intronic splicing silencer. 
     
     
         14 . The method of  claim 12 , wherein the cis splicing regulatory element is an intronic splicing silencer. 
     
     
         15 . The method of  claim 9 , wherein the nucleotide sequence of the antisense oligonucleotide is complementary to a continuous portion of a sequence of intron 6, intron 7, or exon 7 of a survival motor neuron (SMN) 2 gene. 
     
     
         16 . The method of  claim 9 , wherein the nucleotide sequence of the antisense oligonucleotide is complementary to a non-continuous portion of a sequence of intron 6, intron 7, or exon 7 of a survival motor neuron (SMN) 2 gene. 
     
     
         17 . A composition for promoting the incorporation of exon 7 of the SMN2 gene into transcripts of the SMN2 gene, the composition comprising:
 at least a first antisense oligonucleotide having a nucleotide sequence:
 (i) that is chosen from [SEQ ID NO 1], [SEQ ID NO 2], [SEQ ID NO 4], [SEQ ID NO 5], [SEQ ID NO 6], [SEQ ID NO 7], [SEQ ID NO 8], [SEQ ID NO 9], [SEQ ID NO 10], [SEQ ID NO 11], [SEQ ID NO 12], [SEQ ID NO 13], [SEQ ID NO 14], [SEQ ID NO 15], [SEQ ID NO 16], and [SEQ ID NO 17]; or 
 (ii) which is sufficiently homologous to [SEQ ID NO 1], [SEQ ID NO 2], [SEQ ID NO 4], [SEQ ID NO 5], [SEQ ID NO 6], [SEQ ID NO 7], [SEQ ID NO 8], [SEQ ID NO 9], [SEQ ID NO 10], [SEQ ID NO 11], [SEQ ID NO 12], [SEQ ID NO 13], [SEQ ID NO 14], [SEQ ID NO 15], [SEQ ID NO 16], or [SEQ ID NO 17]; 
   such that the nucleotide sequence of the antisense oligonucleotide is complementary to a portion of a sequence of intron 6, intron 7, or exon 7 of the SMN2 gene.   
     
     
         18 . The composition of  claim 17 , further comprising a second antisense oligonucleotide. 
     
     
         19 . The composition of  claim 18 , wherein the first antisense oligonucleotide and the second antisense oligonucleotide are each targeted to the same nucleic acid target. 
     
     
         20 . The composition of  claim 18 , wherein the first antisense oligonucleotide and the second antisense oligonucleotide are each targeted to different nucleic acid targets.