Modulation of transthyretin expression

Abstract

Compounds, compositions and methods are provided for modulating the expression of transthyretin. The compositions comprise oligonucleotides, targeted to nucleic acid encoding transthyretin. Methods of using these compounds for modulation of transthyretin expression and for diagnosis and treatment of diseases and conditions associated with expression of transthyretin are provided.

1 - 18 . (canceled) 
     
     
         19 . A compound comprising a modified oligonucleotide 15 to 30 linked nucleosides in length and having a nucleobase sequence that is at least 95% complementary to SEQ ID NO: 4, and that includes at least 8 consecutive nucleobases of SEQ ID NO: 75;
 wherein each thymine (T) of SEQ ID NO: 75 is replaced by uracil (U);   wherein at least one nucleoside of the modified oligonucleotide comprises a modified sugar moiety;   wherein the compound comprises two deoxythymidine nucleotides (dTdT) attached to the 3′-end of the modified oligonucleotide; and   wherein the compound inhibits the expression of human transthyretin mRNA in cells treated with 50 nM of modified oligonucleotide by at least ten percent (10%), as measured by quantitative real-time PCR.   
     
     
         20 . The compound of  claim 19 , wherein the modified oligonucleotide is an RNA oligonucleotide. 
     
     
         21 . The compound of  claim 20 , comprising a single-stranded RNA oligonucleotide. 
     
     
         22 . The compound of  claim 20 , comprising a double-stranded RNA oligonucleotide. 
     
     
         23 . The compound of  claim 19 , comprising a chimeric oligonucleotide. 
     
     
         24 . The compound of  claim 19 , wherein at least a portion of said compound hybridizes with a target RNA to form an oligonucleotide-target RNA duplex. 
     
     
         25 . The compound of  claim 19  having at least one modified internucleoside linkage, or nucleobase. 
     
     
         26 . The compound of  claim 19  having at least one 2′-O-methoxyethyl sugar moiety. 
     
     
         27 . The compound of  claim 19  having at least one 2′-O-methoxy sugar moiety. 
     
     
         28 . The compound of  claim 19  having at least one 2′-fluoro sugar moiety. 
     
     
         29 . The compound of  claim 24  having at least one phosphorothioate internucleoside linkage. 
     
     
         30 . The compound of  claim 19  wherein at least one cytosine is a 5-methylcytosine. 
     
     
         31 . The compound of  claim 19  wherein the modified oligonucleotide is 100% complementary to SEQ ID NO: 4. 
     
     
         32 . The compound of  claim 19  comprising a conjugate group. 
     
     
         33 . A composition comprising a compound comprising a modified oligonucleotide 15 to 30 linked nucleosides in length and having a nucleobase sequence that is at least 95% complementary to SEQ ID NO: 4, and that includes at least 8 consecutive nucleobases of SEQ ID NO: 75, or a salt thereof, and a pharmaceutically acceptable carrier or diluent;
 wherein each thymine (T) of SEQ ID NO: 75 is replaced by uracil (U);   wherein at least one nucleoside of the modified oligonucleotide comprises a modified sugar moiety;   wherein the compound comprises two deoxythymidine nucleotides (dTdT) attached to the 3′-end of the modified oligonucleotide; and   wherein the compound inhibits the expression of human transthyretin mRNA in cells treated with 50 nM of modified oligonucleotide by at least ten percent (10%), as measured by quantitative real-time PCR.   
     
     
         34 . The composition of  claim 33  wherein the compound comprises a conjugate group. 
     
     
         35 . The composition of  claim 33  wherein the modified oligonucleotide is 100% complementary to SEQ ID NO: 4. 
     
     
         36 . A method of reducing the expression of human transthyretin in a cell or tissue comprising contacting said cell or tissue with composition comprising a compound comprising a modified oligonucleotide 15 to 30 linked nucleosides in length and having a nucleobase sequence that is at least 95% complementary to SEQ ID NO: 4, and that includes at least 8 consecutive nucleobases of SEQ ID NO: 75, or a salt thereof, and a pharmaceutically acceptable carrier or diluent;
 wherein each thymine (T) of SEQ ID NO: 75 is replaced by uracil (U);   wherein at least one nucleoside of the modified oligonucleotide comprises a modified sugar moiety; and   wherein the compound comprises two deoxythymidine nucleotides (dTdT) attached to the 3′-end of the modified oligonucleotide;   wherein expression of human transthyretin is reduced by at least ten percent (10%).   
     
     
         37 . The method of  claim 36  wherein the compound comprises a conjugate group. 
     
     
         38 . The method of  claim 36  wherein the compound is encapsulated within a liposome. 
     
     
         39 . The method of  claim 36  wherein the modified oligonucleotide is 100% complementary to SEQ ID NO: 4.