Compound and method for treating myotonic dystrophy

Abstract

An antisense compound for use in treating myotonic dystrophy DM1 or DM2, a method of enhancing antisense targeting to heart and quadricep muscles, and a method for treating DM1 or DM2 in a mammalian subject are disclosed. The oligonucleotide has 8-30 bases, with at least 8 contiguous bases being complementary to the polyCUG or polyCCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1 or DM2, respectively. Conjugated to the oligonucleotide is a cell-penetrating peptide having the sequence (RXRR(B/X)R) 2 XB, where R is arginine; B is β-alanine; and each X is —C(O)—(CH 2 ) n —NH—, where n is 4-6. The antisense compound is effective to selectively block the sequestration of muscleblind-like 1 protein (MBNL1) and/or CUGBP, in heart and quadricep muscle in a myotonic dystrophy animal model.

1 . An antisense compound for use in treating myotonic dystrophy DM1 or DM2, comprising
 an antisense oligonucleotide having 8-30 bases, with at least 8 contiguous bases being complementary to the polyCUG or polyCCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1 or DM2, respectively, and   conjugated to the oligonucleotide, a cell-penetrating peptide having the sequence (RXRR(B/X)R) 2 XB, where R is arginine; B is β-alanine; and each X is —C(O)—(CH 2 ) n —NH—, where n is 4-6.   where the compound is effective to selectively block the sequestration of at least one of muscleblind-like 1 protein (MBNL1) and CUGBP in heart and quadricep muscle in a myotonic dystrophy animal model.   
     
     
         2 . The antisense compound of  claim 1 , wherein the cell penetrating peptide has the form (RXRRBR) 2 XB and X is —C(O)—(CH 2 ) 6 —NH—, 
     
     
         3 . The antisense compound of  claim 2 , wherein the oligonucleotide is a phosphorodiamidate oligonucleotide (PMO) having between 12-30 bases, and at least 12 contiguous bases that are complementary to the polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1. 
     
     
         4 . The antisense compound of  claim 2 ,  2 , wherein the oligonucleotide is a phosphorodiamidate oligonucleotide (PMO) having between 12-30 bases, and at least 12 contiguous bases that are complementary to the polyCCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM2. 
     
     
         5 . The antisense compound of  claim 1 , wherein the cell penetrating peptide has the form (RXRRXR) 2 XB and X is —C(O)—(CH 2 ) 6 —NH—. 
     
     
         6 . The antisense compound of  claim 5 , wherein the oligonucleotide is a phosphorodiamidate oligonucleotide (PMO) having between 12-30 bases, and at least 12 contiguous bases that are complementary to the polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1. 
     
     
         7 . The antisense compound of  claim 5 , wherein the oligonucleotide is a phosphorodiamidate oligonucleotide (PMO) having between 12-30 bases, and at least 12 contiguous bases that are complementary to the polyCCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM2. 
     
     
         8 . A method of targeting a systemically administered antisense oligonucleotide to heart muscle tissue in a mammalian subject, where the oligonucleotide is directed against the polyCUG or polyCCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1 or DM2, respectively, comprising
 conjugating to the oligonucleotide, a cell-penetrating peptide having the sequence (RXRR(B/X)R) 2 XB, where R is arginine; B is β-alanine; and each X is independently a neutral linear amino acid —C(O)—(CH 2 ) n —NH—, where n is 4-6.   
     
     
         9 . The method of  claim 8 , wherein the selective targeting of the oligonucleotide to heart and quadracep muscle is evidenced by the reversal of sequestration of muscleblind-like 1 protein (MBNL1) in the heart and quadricep muscle tissue in a PMO-treated animal model having known MBNL1 sequestration. 
     
     
         10 . The method of  claim 8 , wherein the cell penetrating peptide has the form (RXRRBR) 2 XB and X is —C(O)—(CH 2 ) 6 —NH—. 
     
     
         11 . The method of  claim 10 , wherein the oligonucleotide is a phosphorodiamidate oligonucleotide (PMO) having between 12-30 bases, and at least 12 contiguous bases that are complementary to (i) the polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1, or (ii) the polyCCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM2. 
     
     
         12 . The method of  claim 8 , wherein the cell penetrating peptide has the form (RXRRXR) 2 XB and X is —C(O)—(CH 2 ) 6 —NH—. 
     
     
         13 . The method of  claim 12 , wherein the oligonucleotide is a phosphorodiamidate oligonucleotide (PMO) having between 12-30 bases, and at least 12 contiguous bases that are complementary to (i) the polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1, or (ii) the polyCCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM2. 
     
     
         14 . A method of treating mytonic dystrophy DM1 or DM2 in a mammalian subject, comprising
 administering to the subject, an antisense compound comprising an antisense oligonucleotide having 8-30 bases, with at least 8 contiguous bases being complementary to the polyCUG or polyCCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1 or DM2, respectively, and conjugated to the oligonucleotide, a cell-penetrating peptide having the sequence (RXRR(B/X)R) 2 XB, where R is arginine; B is β-alanine; and each X is —C(O)—(CH 2 ) n —NH—, where n is 4-6, and   repeating said administering at least once every one week to 3 months.   
     
     
         15 . The method of  claim 14 , for treating DM1 in a mammalian subject, wherein the cell penetrating peptide has the form (RXRRBR) 2 XB, X is —C(O)—(CH 2 ) 6 —NH—, and the oligonucleotide is a phosphorodiamidate oligonucleotide (PMO) having between 12-30 bases, and at least 12 contiguous bases that are complementary to the polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1. 
     
     
         16 . The method of  claim 14 , for treating DM2 in a mammalian subject, wherein the cell penetrating peptide has the form (RXRRBR) 2 XB, X is —C(O)—(CH 2 ) 6 —NH—, and the oligonucleotide is a phosphorodiamidate oligonucleotide (PMO) having between 12-30 bases, and at least 12 contiguous bases that are complementary to the polyCCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM2. 
     
     
         17 . The method of  claim 14 , for treating DM1 in a mammalian subject, wherein the cell penetrating peptide has the form (RXRRXR) 2 XB, X is —C(O)—(CH 2 ) 6 —NH—, and the oligonucleotide is a phosphorodiamidate oligonucleotide (PMO) having between 12-30 bases, and at least 12 contiguous bases that are complementary to the polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1. 
     
     
         18 . The method of  claim 14 , for treating DM2 in a mammalian subject, wherein the cell penetrating peptide has the form (RXRRXR) 2 XB, X is —C(O)—(CH 2 ) 6 —NH—, and the oligonucleotide is a phosphorodiamidate oligonucleotide (PMO) having between 12-30 bases, and at least 12 contiguous bases that are complementary to the polyCCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM2. 
     
     
         19 . The method of  claim 14 , wherein said administering is by intravenous or subcutaneous injection to the subject, at a dose between 1-5 mg/kg body weight antisense compound. 
     
     
         20 . The method of  claim 14 , wherein said administering is repeated repeating step is continued at regular intervals of every one to three months, and further includes monitoring the patient during the treatment period for improvement in skeletal or heart muscle performance. 
     
     
         21 . The method of  claim 14 , wherein said administering is repeated repeating step is continued at regular intervals of every one to three months, and further includes monitoring the patient during the treatment period for improvement in heart conduction properties. 
     
     
         22 . The method of  claim 20 , wherein said administering is repeated repeating step is continued at regular intervals of every one to three months, and further includes monitoring the patient during the treatment period for reduction in serum creatine kinase.