Compositions for Modulating C9ORF72 Expression

Abstract

Disclosed herein are compositions and methods for reducing expression of C9ORF72 mRNA and protein in an animal with C9ORF72 specific inhibitors. Also disclosed herein are compositions and methods of selectively inhibiting a C9ORF72 pathogenic associated mRNA variant by administering an antisense compound targeting the region beginning at the start site of exon 1A to the start site of exon 1B of a C9ORF72 pre-mRNA. Such methods are useful to treat, prevent, or ameliorate neurodegenerative diseases in an individual in need thereof. Such C9ORF72 specific inhibitors include antisense compounds.

1 . A compound, comprising a modified oligonucleotide consisting of 12 to 50 linked nucleosides, wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to an equal length portion of a C9ORF72 nucleic acid, and wherein the modified oligonucleotide comprises at least one modification selected from a modified sugar, a sugar surrogate, and a modified internucleoside linkage. 
     
     
         2 . A compound, comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 12 consecutive nucleobases of any of the nucleobase sequences of SEQ ID NOs: 20-401, 441, 468-554, 591-1053, 1055-1318, 1366-1488, and 1492-1545. 
     
     
         3 . A compound comprising a modified oligonucleotide consisting of 12 to 30 linked nucleosides and having a nucleobase sequence comprising at least 8 consecutive nucleobases complementary to an equal length portion of nucleobases 1107-1520, 1111-1200, 1211-1318, 1326-1540, 1331-1375, 1368-1391, 1398-1424, 1411-1440, 1429-1481, 1502-1539, 1508-1539, 7989-8038, 8020-8135, 8136-8161, or 8174-8211 of SEQ ID NO: 2. 
     
     
         4 . The compound of  claim 3 , wherein the modified oligonucleotide is a single-stranded modified oligonucleotide. 
     
     
         5 . The compound of  claim 3 , wherein at least one internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage. 
     
     
         6 . The compound of  claim 5 , wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage. 
     
     
         7 . The compound of  claim 5 , wherein each internucleoside linkage of the modified oligonucleotide is a modified internucleoside linkage. 
     
     
         8 . The compound of  claim 7 , wherein the modified internucleoside linkage is a phosphorothioate internucleoside linkage. 
     
     
         9 . The compound of  claim 3 , wherein at least one internucleoside linkage of the modified oligonucleotide is a phosphodiester internucleoside linkage. 
     
     
         10 . The compound of  claim 3 , wherein at least one nucleobase of the modified oligonucleotide comprises a modified nucleobase. 
     
     
         11 . The compound of  claim 10 , wherein the modified nucleobase is a 5-methylcytosine. 
     
     
         12 . The compound of  claim 3 , wherein at least one nucleoside of the modified oligonucleotide comprises a modified sugar. 
     
     
         13 . The compound of  claim 12 , wherein each nucleoside of the modified oligonucleotide comprises a modified sugar. 
     
     
         14 . The compound of  claim 12 , wherein the modified sugar is a bicyclic sugar. 
     
     
         15 . The compound of  claim 14 , wherein each bicyclic sugar comprises a chemical bridge between the 4′ and 2′ positions of the sugar, wherein each chemical bridge is independently selected from: 4′-CH(R)—O-2′ and 4′-(CH 2 ) 2 —O-2′, wherein R is independently selected from H, C 1 -C 6  alkyl, and C 1 -C 6  alkoxy. 
     
     
         16 . The compound of  claim 15 , wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is methyl. 
     
     
         17 . The compound of  claim 15 , wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is H. 
     
     
         18 . The compound of  claim 15 , wherein at least one chemical bridge is 4′-CH(R)—O-2′ and wherein R is —CH 2 —O—CH 3 . 
     
     
         19 . The compound of  claim 12 , wherein at least one modified sugar comprises a 2′-O-methoxyethyl group. 
     
     
         20 . The compound of  claim 12 , wherein at least one modified sugar comprises a 2′-O-methyl group. 
     
     
         21 . The compound of  claim 13 , wherein each modified sugar comprises a 2′-O-methoxyethyl group or a 2′-O-methyl group. 
     
     
         22 . The compound of  claim 3 , wherein the modified oligonucleotide is a gapmer. 
     
     
         23 . The compound of  claim 3 , wherein the modified oligonucleotide comprises:
 a gap segment consisting of 10 linked deoxynucleosides;   a 5′ wing segment consisting of 5 linked nucleosides; and   a 3′ wing segment consisting of 5 linked nucleosides;   wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.   
     
     
         24 . The compound of  claim 3 , wherein the modified oligonucleotide comprises:
 a gap segment consisting of 8 linked deoxynucleosides;   a 5′ wing segment consisting of 5 linked nucleosides; and   a 3′ wing segment consisting of 5 linked nucleosides;   wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment and wherein each nucleoside of each wing segment comprises a modified sugar.   
     
     
         25 . The compound of  claim 3 , wherein the modified oligonucleotide comprises sugar modifications in any of the following patterns: eeekkdddddddkkeee, eekkddddddddkkeee, ekddddddddekekeee, kekeddddddddekeke, and ekekddddddddkekee; wherein,
 e=a 2′-O-methoxyethyl modified nucleoside,   d=a 2′-deoxynucleoside, and   k=a cEt nucleoside.   
     
     
         26 . The compound of  claim 25 , wherein the modified oligonucleotide comprises internucleoside linkages in any of the following patterns: soooossssssssssooss, sooosssssssssooss, soosssssssssooss, and sosssssssssoooss; wherein,
 s=a phosphorothioate linkage, and   o=a phosphodiester linkage.   
     
     
         27 . The compound of  claim 3 , wherein the modified oligonucleotide consists of 17, 18, 19, or 20 linked nucleosides. 
     
     
         28 .- 30 . (canceled) 
     
     
         31 . The compound of  claim 3 , wherein the modified oligonucleotide has a nucleobase sequence complementary to a region of C9ORF72 other than a hexanucleotide repeat expansion, wherein the hexanucleotide repeat expansion comprises any of GGGCC, GGGGGG, GGGGCG, and GGGGGC. 
     
     
         32 . A conjugated antisense compound comprising the compound of  claim 3 . 
     
     
         33 . The compound of  claim 3 , consisting of the modified oligonucleotide. 
     
     
         34 . A double-stranded compound comprising the compound of  claim 3 . 
     
     
         35 . A conjugated antisense compound comprising the double-stranded compound of  claim 34 . 
     
     
         36 . A composition comprising a compound according to  claim 3  and at least one of a pharmaceutically acceptable carrier or diluent, wherein optionally the pharmaceutically acceptable diluent is phosphate-buffered saline (PBS). 
     
     
         37 . The composition according to  claim 36 , wherein the modified oligonucleotide of the compound is a salt, wherein optionally the salt is a sodium salt. 
     
     
         38 . (canceled) 
     
     
         39 . (canceled)