Compound and method for treating myotonic dystrophy

• US 8,741,863 B2
• Filed: 08/26/2011
• Issued: 06/03/2014
• Est. Priority Date: 06/29/2007
• Status: Active Grant

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: Moulton, Hong M., Kole, Ryszard
• Primary Examiner(s): VIVLEMORE, TRACY ANN
• Application Number:
  US13/219,401
• Publication Number:
  US 20120058946A1
• Time in Patent Office: 1,012 Days
• Field of Search: None
• US Class Current: 514/44.A
• CPC Class Codes: A61K 31/7088 Compounds having three or more nucleosides or nucleotides
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C07K 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2810/40 Vectors comprising a peptide as targeting moiety, e.g. a synthetic peptide, from undefined source

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)₂XB, where R is arginine; B is β-alanine; and each X is —C(O)—(CH₂)_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.

Claims

1. 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 (RXRRBR)₂XB, where R is arginine;

B is β

-alanine; and

each X is independently —

C(O)—

(CH₂)_n—

NH—

, where n is 4-6 (SEQ ID NO;

     67),

where the compound is effective to selectively block the sequestration of at least one of muscleblind-like 1 protein (MBNL1) and CUG-binding protein 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 sequence (RXRRBR)₂XB (SEQ ID NO:

19) where X is —

C(O)—

(CH₂)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, 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. 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, the method comprising

conjugating to the oligonucleotide, a cell-penetrating peptide having the sequence (RXRRBR)₂XB, where R is arginine;

B is β

-alanine; and

each X is independently —

C(O)—

(CH₂)_n—

NH—

, where n is 4-6 (SEQ ID NO;

     67).

6. The method of claim 5, wherein the selective targeting of the oligonucleotide to heart muscle tissue is evidenced by the reversal of sequestration of muscleblind-like 1 protein (MBNL1) in the heart muscle tissue in a PMO-treated animal model having known MBNL1 sequestration.

7. The method of claim 5, wherein the cell penetrating peptide has the sequence(RXRRBR)₂XB (SEQ ID NO:

19) where X is —

C(O)—

(CH₂)₆—

NH—

.

8. The method of claim 7, 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.

9. A method of treating mytonic dystrophy DM1 or DM2 in a mammalian subject, the method 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 RXRRBR)₂XB, where R is arginine;

B is β

-alanine; and

each X is independently —

C(O)—

(CH₂)_n—

NH—

, where n is 4-6 (SEQ ID NO;

     67), and

repeating said administering at least once every one week to 3 months.

10. The method of claim 9, for treating DM1 in a mammalian subject, wherein the cell penetrating peptide has the form (RXRRBR)₂XB (SEQ ID NO:

19), where X is —

C(O)—

(CH₂)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.

11. The method of claim 9, for treating DM2 in a mammalian subject, wherein the cell penetrating peptide has the form (RXRRBR)₂XB (SEQ ID NO:

19), where X is —

C(O)—

(CH₂)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.

12. The method of claim 9, wherein said administering is by intravenous or subcutaneous injection to the subject, at a dose between 1-5 mg/kg body weight antisense compound.

13. The method of claim 9, wherein said administering is repeated 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.

14. The method of claim 9, wherein said administering is repeated 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.

15. The method of claim 9, wherein said administering is repeated 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.