Oligonucleotides

Abstract

The present invention relates to oligonucleotides that maintain a Toll-Like Receptor 7 (TLR7) response and/or which potentiate Toll-Like Receptor 8 (TLR8) sensing.

1 - 62 . (canceled) 
     
     
         63 . An oligonucleotide comprising one or more modified bases and at least four thymidines, wherein the oligonucleotide potentiates Toll-like receptor 8 (TLR8) activity when administered to an animal. 
     
     
         64 . The oligonucleotide of  claim 63  which comprises
 a) a 5′ region at least five bases in length which are modified and/or which have a modified backbone, 
 b) a middle region comprising a stretch of ten bases, wherein at least four of the bases are thymidine, 
 c) a 3′ region at least five bases in length. 
 
     
     
         65 . The oligonucleotide of  claim 63 , wherein
 i) the at least four thymidine bases are in a continuous stretch, and/or   
       ii) one, two, three or four of the at least four thymidine bases are not in a continuous stretch. 
     
     
         66 - 68 . (canceled) 
     
     
         69 . The oligonucleotide according to  claim 63 , comprising, consisting of or consisting essentially of a nucleic acid sequence set forth in Tables 1 to 6 or a variant thereof. 
     
     
         70 - 72 . (canceled) 
     
     
         73 . A method of reducing expression of a target gene in a cell, the method comprising contacting the cell with an oligonucleotide according to  claim 63 . 
     
     
         74 . A method of treating or preventing a disease in a subject, the method comprising administering to the subject an oligonucleotide according to  claim 63 , wherein the oligonucleotide reduces the expression of a target gene involved in the disease. 
     
     
         75 . The method of  claim 74 , wherein the animal has been, or will be, administered with an immune response modifier. 
     
     
         76 . The method of  claim 75 , wherein the immune response modifier is a Toll-like receptor (TLR) agonist such as a base analog (including: a guanosine analog, a deaza-adenosine analog, an imidazoquinoline or a derivative, a hydroxyadenine compound or a derivative, a thiazoloquinolone compound or a derivative, a benzoazepine compound or a derivative), or an RNA molecule. 
     
     
         77 - 80 . (canceled) 
     
     
         81 . The oligonucleotide according to  claim 63 , comprising a 5′ region, a 3′ region and a middle region, comprising ribonucleic acid, or deoxyribonucleic acid bases or a combination thereof, wherein one or both of the 5′ region and the 3′ region comprise bases which are modified and/or which have a modified backbone, and at least one of the following apply;
 a) the 5′ region comprises three continuous pyrimidine bases which are modified and/or which have a modified backbone, 
 b) the 5′ region comprises bases which are modified and/or which have a modified backbone and the junction between the 5′ region and middle region comprises three continuous pyrimidine bases, 
 c) the 3′ region comprises three continuous pyrimidine bases which are modified and/or which have a modified backbone, 
 d) the 3′ region comprises bases which are modified and/or which have a modified backbone and the junction between the 3′ region and middle region comprises three continuous pyrimidine bases, and 
 e) the 5′ region comprises two continuous cytosine bases which are modified and/or which have a modified backbone. 
 
     
     
         82 . The oligonucleotide according to  claim 81 , wherein the three continuous pyrimidine bases are within seven bases of the 5′ and/or 3′ end of the oligonucleotide. 
     
     
         83 . The oligonucleotide according to  claim 81 , wherein the three continuous pyrimidine bases are at the 5′ and/or 3′ end of the oligonucleotide. 
     
     
         84 . The oligonucleotide according to  claim 81 ,
 wherein the 5′ three continuous pyrimidine bases have the sequence 5′-CUU-3′, 5′-CUT-3′, 5′-CCU-3′, 5′-UUC-3′, 5′-UUU-3′ or 5′-CTT-3′; or   wherein the 3′three continuous pyrimidine bases have the sequence 5′-UUC-3′, 5′-TUC-3′, 5′-UCC-3′, 5′-CUU-3′, 5′-UUU-3′ or 5′-TTC-3′.   
     
     
         85 . The oligonucleotide according to  claim 81 , wherein the two continuous cytosine bases comprise a 2′-LNA and a phosphorothioate backbone. 
     
     
         86 . The oligonucleotide according to  claim 81 , wherein at least one of the three continuous pyrimidine bases and/or at least one of the two continuous cytosine bases does not hybridize to a target polynucleotide. 
     
     
         87 . The oligonucleotide according to  claim 63 , wherein the modified base comprises a 2′-O-methyl, 2′-O-methoxyethoxy, 2′-fluoro, 2′-allyl, 2′-O-[2-(methylamino)-2-oxoethyl], 4′-thio, 4′-CH2-O-T-bridge, 4′-(CH2)2-O-2′-bridge, 2′-LNA, 2′-amino, fluoroarabinonucleotide, threose nucleic acid or 2′-O—(N-methlycarbamate). 
     
     
         88 . The oligonucleotide according to  claim 63 , wherein the modified backbone comprises a phosphorothioate, a non-bridging oxygen atom substituting a sulfur atom, a phosphonate such as a methylphosphonate, a phosphodiester, a phosphoromorpholidate, a phosphoropiperazidate, amides, methylene(methylamino), fromacetal, thioformacetal, a peptide nucleic acid or a phosphoroamidate such as a morpholino phosphorodiamidate (PMO), N3′-P5′ phosphoramidite or thiophosphoroamidite. 
     
     
         89 . The oligonucleotide according to  claim 63 , wherein the modified base comprises a 2′O-methyl and a phosphorothioate backbone. 
     
     
         90 . The oligonucleotide according to  claim 63 , wherein the oligonucleotide is at least 10, at least about 18, at least about 20, at about least 25, between about 10 and about 50 nucleotides, between about 18 and about 50 nucleotides, between about 18 and about 30 nucleotides, between about 20 and about 30 nucleotides, between about 20 and 5,000 nucleotides, or about 20 bases in length. 
     
     
         91 . The oligonucleotide according to  claim 63 , wherein the oligonucleotide is an antisense oligonucleotide or a double stranded oligonucleotide for gene silencing. 
     
     
         92 . The oligonucleotide according to  claim 63 , wherein one, two or all three of the three continuous pyrimidine bases are removed by an endonuclease in vivo.