Pharmaceutical Composition

Abstract

The invention provides pharmaceutical compositions comprising short single stranded oligonucleotides, of length of between 8 and 17 nucleobases which are complementary to human microRNAs. The short oligonucleotides are particularly effective at alleviating miRNA repression in vivo. It is found that the incorporation of high affinity nucleotide analogues into the oligonucleotides results in highly effective anti-microRNA molecules which appear to function via the formation of almost irreversible duplexes with the miRNA target, rather than RNA cleavage based mechanisms, such as mechanisms associated with RNaseH or RISC.

1 - 95 . (canceled) 
     
     
         96 . An oligonucleotide 8-22 nucleotides in length, for inhibition of a microRNA target in a cell, wherein the oligonucleotide comprises Locked Nucleic Acid (LNA) nucleobase units; wherein at least 30% of the nucleobases are LNA units; wherein the oligonucleotide does not comprise a region of more than 5 consecutive 2′ deoxyribose nucleoside units; wherein the oligonucleotide does not comprise a region of more than 2 consecutive LNA units; and wherein said oligonucleotide comprises at least one phosphorothioate internucleoside linkage group. 
     
     
         97 . The oligonucleotide according to  claim 96 , wherein all the internucleoside linkage groups are phosphorothioate linkages. 
     
     
         98 . The oligonucleotide according to  claim 96 , wherein the LNA units are beta D oxy-LNA units. 
     
     
         99 . The oligonucleotide according to  claim 98  wherein the first nucleobase of the oligonucleotide, counting from the 3′ end, is an LNA unit. 
     
     
         100 . The oligonucleotide according to 99 wherein the second nucleobase of the oligonucleotide, counting from the 3′ end, is an LNA unit. 
     
     
         101 . The oligonucleotide according to  claim 96 , wherein the oligonucleotide comprises at least one region of two consecutive LNA units. 
     
     
         102 . The oligonucleotide according to  claim 96 , wherein the oligonucleotide comprises an LNA unit at the 5′ end. 
     
     
         103 . The oligonucleotide according to  claim 96 , wherein at least 50% of the nucleobase units of the oligonucleotide are LNA nucleobase units. 
     
     
         104 . The oligonucleotide according to  claim 96  wherein the oligonucleotide comprises at least one further non-LNA nucleotide analogue unit. 
     
     
         105 . The oligonucleotide according to  claim 104  wherein said at least one further non-LNA nucleotide analogue unit is a DNA nucleotide analogue where the 2′ hydrogen group of the DNA nucleotide has been substituted with a group selected from the group consisting of —O—CH 3 , O—CH 2 —CH 2 —O—CH 3 , —O—CH 2 —CH 2 —CH 2 —NH 2 , O—CH 2 —CH 2 —CH 2 —OH and —F. 
     
     
         106 . The oligonucleotide according to  claim 104 , wherein the non-LNA nucleotide analogue units are 2′-MOE nucleotide analogues. 
     
     
         107 . The oligonucleotide according to  claim 104 , wherein the non-LNA nucleotide analogue unit or units are independently selected from 2′-OMe RNA units and 2′-fluoro DNA units. 
     
     
         108 . The oligonucleotide according to  claim 96  wherein each nucleobase unit is independently selected from the group consisting of LNA nucleobase units and DNA nucleobase units. 
     
     
         109 . The oligonucleotide according to  claim 96 , wherein the oligonucleotide comprises 5 LNA units. 
     
     
         110 . The oligonucleotide according to  claim 96 , wherein at least 3 of the LNA nucleobases are either cytosine or guanine. 
     
     
         111 . The oligonucleotide according to  claim 96 , wherein the oligonucleotide comprises both LNA nucleotide analogue units and non-LNA nucleotide analogue units and all the nucleobase units of the oligonucleotide are nucleotide analogues. 
     
     
         112 . The oligonucleotide according to 96, wherein the oligonucleotide has a length of 10 to 16 nucleobases. 
     
     
         113 . The oligonucleotide according to 96, wherein the oligonucleotide has a length of 15 or 16 nucleobases. 
     
     
         114 . The oligonucleotide according to  claim 96 , wherein the oligonucleotide has a length of 15 nucleotides and the substitution pattern, starting from the 3′ end, is 3′ XXxXxXxxXXxxXxX 5′, wherein “X” denotes an LNA unit and “x” denotes a DNA unit.