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 for the inhibition of microRNA-155 having a length of 12-16 nucleotides, wherein:
 (a) said oligonucleotide is fully complementary to a human microRNA-155 sequence;   (b) a first and a second nucleotide from a 3′ end of the oligonucleotide are LNAs;   (c) the oligonucleotide does not comprise a region of more than 5 consecutive DNA units; and   (d) the oligonucleotide comprises at least one phosphorothioate internucleotide linkage.   
     
     
         97 . The oligonucleotide of  claim 96 , wherein the oligonucleotide has a length of 12, 13, 14, 15, or 16 nucleotides. 
     
     
         98 . The oligonucleotide of  claim 96 , wherein a ninth and/or a tenth nucleotide from the 3′ end of the oligonucleotide is an LNA. 
     
     
         99 . The oligonucleotide of  claim 96 , wherein the oligonucleotide does not comprise a region of more than 3 consecutive DNA units. 
     
     
         100 . The oligonucleotide of  claim 96 , wherein all internucleotide linkages are phosphorothioate linkages. 
     
     
         101 . The oligonucleotide of  claim 96 , wherein a sequence of the first three nucleotides from a 5′ end of the oligonucleotide is selected from the group consisting of Xxx, xXx, xxX, XXx, XxX, xXX and XXX, wherein “X” denotes a LNA unit, and “x” denotes a DNA unit. 
     
     
         102 . The oligonucleotide of  claim 96 , wherein the oligonucleotide comprises at least three LNA units at positions 1 to 7 from the 3′ end of the oligonucleotide. 
     
     
         103 . The oligonucleotide of  claim 96 , wherein the oligonucleotide comprises at least four LNA units at positions 1 to 7 from the 3′ end of the oligonucleotide. 
     
     
         104 . The oligonucleotide of  claim 96 , wherein the oligonucleotide comprises at least five LNA units at positions 1 to 7 from the 3′ end of the oligonucleotide. 
     
     
         105 . The oligonucleotide of  claim 96 , wherein the oligonucleotide has a length of 14 nucleotides and a sequence of four nucleotides at positions 11 to 14 from the 3′ end of the oligonucleotide is selected from the group consisting of xxxx, Xxxx, xXxx, xxXx, xxxX, XXxx, XxXx, XxxX, xXXx, xXxX, xxXX, XXXx, XxXX, xXXX, XXxX and XXXX, wherein X is an LNA unit and x is a DNA or RNA nucleotide unit. 
     
     
         106 . The oligonucleotide of  claim 96 , wherein the oligonucleotide has a length of 13 nucleotides and a sequence of three nucleotides at positions 11 to 13 from the 3′ end of the oligonucleotide is selected from the group consisting of xxx, Xxx, xXx, xxX, XXx, XxX, xXX and XXX, wherein X is an LNA unit and x is a DNA or RNA nucleotide unit. 
     
     
         107 . The oligonucleotide of  claim 96 , wherein the oligonucleotide has a length of 12 nucleotides and a sequence of two nucleotides at positions 11 to 12 from the 3′ end of the oligonucleotide is selected from the group consisting of xx, XX, xX and Xx, wherein X is an LNA unit and x is a DNA or RNA nucleotide unit. 
     
     
         108 . The oligonucleotide of  claim 96 , wherein the oligonucleotide has a length of 15 nucleotides and a sequence of five nucleotides at positions 11 to 15 from the 3′ end of the oligonucleotide is selected from the group consisting of Xxxxx, xXxxx, xxXxx, xxxXx, xxxxX, XXxxx, XxXxx, XxxXx, XxxxX, xXXxx, xXxXx, xXxxX, xxXXx, xxXxX, xxxXX, XXXxx, XXxxX, XxxXX, xXXXx, xxXXX, XXxXX, XxXxX, XXXXx, XXXxX, XXxXX, XxXXXX, xXXXX, and XXXXX, wherein X is an LNA unit and x is a DNA or RNA nucleotide unit. 
     
     
         109 . The oligonucleotide of  claim 96 , wherein the oligonucleotide has a length of 16 nucleotides and a sequence of six nucleotides at positions 11 to 16 from the 3′ end of the oligonucleotide is selected from the group consisting of Xxxxxx, xXxxxx, xxXxxx, xxxXxx, xxxxXx, xxxxxX, XXxxxx, XxXxxx, XxxXxx, XxxxXx, XxxxxX, xXXxxx, xXxXxx, xXxxXx, xXxxxX, xxXXxx, xxXxXx, xxXxxX, xxxXXx, xxxXxX, xxxxXX, XXXxxx, XXxXxx, XXxxXx, XXxxxX, XxXXxx, XxXxXx, XxXxxX, XxxXXx, XxxXxX, XxxxXX, xXXXxx, xXXxXx, xXXxxX, xXxXXx, xXxXxX, xXxxXX, xxXXXx, xxXXxX, xxXxXX, xxxXXX, XXXXxx, XXXxxX, XXxxXX, XxxXXX, xxXXXX, xXxXXX, XxXxXX, XXxXxX, XXXxXx, xXXxXX, XxXXxX, XXxXXx, xXXXxX, XxXXXx, xXXXXx, xXXXXX, XxXXXX, XXxXXX, XXXxXX, XXXXxX, XXXXXx, and XXXXXX, wherein X is an LNA unit and x is a DNA or RNA nucleotide unit. 
     
     
         110 . The oligonucleotide according to  claim 96 , wherein the LNAs are beta D oxy-LNAs. 
     
     
         111 . A composition comprising the oligonucleotide of  claim 96  and a pharmaceutically acceptable diluent, carrier or adjuvant, or combinations thereof.