Compounds and methods for modulation of dystrophia myotonica-protein kinase (dmpk) expression

Abstract

Provided herein are methods, compounds, and compositions for reducing expression of a DMPK mRNA and protein in an animal. Also provided herein are methods, compounds, and compositions for preferentially reducing CUGexp DMPK RNA, reducing myotonia or reducing spliceopathy in an animal. Such methods, compounds, and compositions are useful to treat, prevent, delay, or ameliorate type 1 myotonic dystrophy, or a symptom thereof.

1 - 167 . (canceled) 
     
     
         168 . A compound comprising a modified oligonucleotide consisting of 10-30 linked nucleosides and having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases 100% complementary to an equal length portion of nucleobases 730-748, 1317-1366, 1343-1368, or 2748-2791 of SEQ ID NO: 1 or nucleobases 8603-8619, 10201-10216, 10202-10218, or 13836-13851 of SEQ ID NO: 2, wherein the nucleobase sequence of the modified oligonucleotide is at least 90% complementary to SEQ ID NO: 1 or SEQ ID NO: 2 as measured over the entirety of the modified oligonucleotide. 
     
     
         169 . The compound of  claim 168 , wherein the modified oligonucleotide is a single-stranded oligonucleotide. 
     
     
         170 . The compound of  claim 169 , wherein at least one nucleoside comprises a modified nucleobase. 
     
     
         171 . The compound of  claim 170 , wherein the modified nucleobase is a 5-methylcytosine. 
     
     
         172 . The compound of  claim 171 , wherein at least one internucleoside linkage is a modified internucleoside linkage. 
     
     
         173 . The compound of  claim 170 , wherein each internucleoside linkage is a phosphorothioate internucleoside linkage. 
     
     
         174 . The compound of  claim 173 , wherein at least one nucleoside comprises a modified sugar. 
     
     
         175 . The compound of  claim 173 , wherein at least two nucleosides comprise a modified sugar. 
     
     
         176 . The compound of  claim 175 , wherein each of the modified sugars have the same modification. 
     
     
         177 . The compound of  claim 175 , wherein at least one of the modified sugars has a different modification. 
     
     
         178 . The compound of  claim 174 , wherein the modified sugar is a bicyclic sugar. 
     
     
         179 . The compound of  claim 178 , wherein the bicyclic sugar is selected from among cEt, LNA, α-L-LNA, ENA, and 2′-thio LNA. 
     
     
         180 . The compound of  claim 174 , wherein the at least one nucleoside comprising a modified sugar is a 2′-substituted nucleoside. 
     
     
         181 . The compound of  claim 180 , wherein the 2′-substituted nucleoside is selected from among: 2′-OCH 3 , 2′-F, and 2′-O-methoxyethyl. 
     
     
         182 . The compound of  claim 169 , wherein the modified oligonucleotide comprises:
 a. a gap segment consisting of linked deoxynucleosides;   b. a 5′ wing segment consisting of liked nucleosides;   c. a 3′ wing segment consisting of linked nucleosides;   d. 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.   
     
     
         183 . The compound of  claim 182 , wherein the modified oligonucleotide consists of 16, 17, 18, 19, or 20 linked nucleosides. 
     
     
         184 . A compound comprising a modified oligonucleotide consisting of 10-30 linked nucleosides and having a nucleobase sequence comprising a portion of at least 8 contiguous nucleobases of any of SEQ ID NOs: 23, 25, 26, 28, 29, 30, 31, and 32. 
     
     
         185 . The compound of  claim 184 , wherein the modified oligonucleotide is a single-stranded oligonucleotide. 
     
     
         186 . The compound of  claim 185 , wherein at least one nucleoside comprises a modified nucleobase. 
     
     
         187 . The compound of  claim 186 , wherein the modified nucleobase is a 5-methylcytosine. 
     
     
         188 . The compound of  claim 187 , wherein at least one internucleoside linkage is a modified internucleoside linkage. 
     
     
         189 . The compound of  claim 188 , wherein each internucleoside linkage is a phosphorothioate internucleoside linkage. 
     
     
         190 . The compound of  claim 189 , wherein at least one nucleoside comprises a modified sugar. 
     
     
         191 . The compound of  claim 189 , wherein at least two nucleosides comprise a modified sugar. 
     
     
         192 . The compound of  claim 191 , wherein each of the modified sugars have the same modification. 
     
     
         193 . The compound of  claim 191 , wherein at least one of the modified sugars has a different modification. 
     
     
         194 . The compound of  claim 190 , wherein the modified sugar is a bicyclic sugar. 
     
     
         195 . The compound of  claim 194 , wherein the bicyclic sugar is selected from among cEt, LNA, α-L-LNA, ENA, and 2′-thio LNA. 
     
     
         196 . The compound of  claim 194 , wherein the at least one nucleoside comprising a modified sugar is a 2′-substituted nucleoside. 
     
     
         197 . The compound of  claim 196 , wherein the 2′-substituted nucleoside is selected from among: 2′-OCH 3 , 2′-F, and 2′-O-methoxyethyl. 
     
     
         198 . The compound of  claim 185 , wherein the modified oligonucleotide comprises:
 a. a gap segment consisting of linked deoxynucleosides;   b. a 5′ wing segment consisting of liked nucleosides;   c. a 3′ wing segment consisting of linked nucleosides;   d. 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.   
     
     
         199 . The compound of  claim 198 , wherein the modified oligonucleotide consists of 16, 17, 18, 19, or 20 linked nucleosides. 
     
     
         200 . A compound comprising a modified oligonucleotide having a nucleobase sequence as set forth in SEQ ID NO: 30, wherein the modified oligonucleotide consists of 16 linked nucleosides and comprises:
 a. a gap segment consisting of eight linked deoxynucleosides;   b. a 5′ wing segment consisting of four linked nucleosides and having a E-E-K-K 5′-wing motif;   c. a 3′ wing segment consisting of four linked nucleosides and having a K-K-E-E 3′-wing motif;   d. wherein the gap segment is positioned between the 5′ wing segment and the 3′ wing segment;   e. wherein each E represents 2′-O-methoxyethyl sugar and each K represents a cEt sugar;   f. wherein each internucleoside linkage is a phosphorothioate internucleoside linkage; and   g. wherein each cytosine residue is a 5-methyl cytosine.   
     
     
         201 . A composition comprising the compound of  claim 168  or a salt thereof and a pharmaceutically acceptable carrier or diluent. 
     
     
         202 . A composition comprising the compound of  claim 184  or a salt thereof and a pharmaceutically acceptable carrier or diluent. 
     
     
         203 . A composition comprising the compound of  claim 200  or a salt thereof and a pharmaceutically acceptable carrier or diluent. 
     
     
         204 . A method of treating DM1 in an animal comprising administering to an animal in need thereof a composition according to  claim 201 . 
     
     
         205 . A method of treating DM1 in an animal comprising administering to an animal in need thereof a composition according to  claim 202 . 
     
     
         206 . A method of treating DM1 in an animal comprising administering to an animal in need thereof a composition according to  claim 203 .