Compound and method for treating myotonic dystrophy

Patent metadata

Field	Value
Publication number	US-10106796-B2
Application number	US-201414464561-A
Country	US
Kind code	B2
Filing date	Aug 20, 2014
Priority date	Jun 26, 2009
Publication date	Oct 23, 2018
Grant date	Oct 23, 2018

Abstract

Provided are 9-base morpholino antisense compounds targeted to polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA, and related methods for treating myotonic dystrophy DM1.

First claim

It is claimed:

1. An antisense compound, comprising
an uncharged antisense oligomer of 8-30 bases comprising a sequence of at least 8 contiguous bases that is complementary to: (i) the polyCUG repeats in the 3′-UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in Myotonic dystrophy type 1 (DM1) or (ii) the polyCCUG repeats in the first intron of zinc finger protein 9 (ZNF9) mRNA in Myotonic dystrophy type 2 (DM2), and
a cell-penetrating peptide (CPP) conjugated to the oligomer, wherein the CPP is of a formula selected from the group consisting of:

wherein independently for each instance R a is H or acetyl.

2. The antisense compound of claim 1 , wherein the CPP is conjugated to the antisense oligomer with a linker selected from glycine (G), cysteine (C), 6-aminohexanoic acid (Ahx), β-alanine (B), and AhxB.

3. The antisense compound of claim 2 , wherein the linker is G.

4. The antisense compound of claim 2 , wherein the CPP and linker together are of a formula selected from the group consisting of:

wherein independently for each instance R a is H or acetyl.

5. The antisense compound of claim 1 , wherein the antisense oligomer is a phosphorodiamidate morpholino oligomer (PMO) having a sequence complementary to the polyCUG repeats in the 3′-UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1.

6. The antisense compound of claim 5 , wherein the sequence is selected from SEQ ID NOs: 1-14.

7. The antisense compound of claim 1 , wherein the antisense oligomer is a phosphorodiamidate morpholino oligomer (PMO) having a sequence complementary to the polyCCUG repeats in the first intron of zinc finger protein 9 (ZNF9) mRNA in DM2.

8. The antisense compound of claim 7 , wherein the sequence is selected from SEQ ID NOs: 19-25.

9. An antisense compound, comprising:
an antisense oligomer of 8-30 morpholino subunits linked together by phosphorodiamidate linkages, wherein:
each morpholino subunit is of a formula:

wherein each B is a base pairing moiety independently selected from adenine, cytosine, guanine, uracil, thymine or hypoxanthine, wherein each B taken together comprises a sequence of at least 8 contiguous bases that is complementary to: (i) the polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in Myotonic dystrophy type 1 (DM1) or (ii) the polyCCUG repeats in the first intron of zinc finger protein 9 (ZNF9) mRNA in Myotonic dystrophy type 2 (DM2); and
each phosphorodiamidate linkage is of a formula:

 and

a cell-penetrating peptide (CPP) conjugated to the morpholino oligomer, wherein:
the CPP is of a formula selected from the group consisting of:

wherein independently for each instance R a is H or acetyl, and the CPP is conjugated to the morpholino oligomer by a linker selected from:

10. The antisense compound of claim 9 , wherein the CPP and linker together are of a formula selected from the group consisting of:

wherein independently for each instance R a is H or acetyl.

11. The antisense compound of claim 9 , wherein the sequence is complementary to the polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in DM1.

12. The antisense compound of claim 11 , wherein the sequence is selected from SEQ ID NOs: 1-14.

13. The antisense compound of claim 9 , wherein the sequence is complementary to the polyCCUG repeats in the first intron of zinc finger protein 9 (ZNF9) mRNA in DM2.

14. The antisense compound of claim 13 , wherein the sequence is selected from SEQ ID NOs: 19-25.

15. A method of treating myotonic dystrophy type 1 (DM1) or myotonic dystrophy type 2 (DM2) in a mammalian subject in need thereof, comprising administering to the subject an effective amount of an antisense compound, wherein the antisense compound comprises:
an antisense oligomer of 8-30 morpholino subunits linked together by phosophorodiamidate linkages, wherein:
each morpholino subunit is of a formula:

wherein each B is a base pairing moiety independently selected from adenine, cytosine, guanine, uracil, thymine or hypoxanthine, wherein each B taken together comprises a sequence of at least 8 contiguous bases that is complementary to: (i) the polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in Myotonic dystrophy type 1 (DM1) or (ii) the polyCCUG repeats in the first intron of zinc finger protein 9 (ZNF9) mRNA in Myotonic dystrophy type 2 (DM2); and

each phosphorodiamidate linkage is of a formula:

 and

a cell-penetrating peptide (CPP) conjugated to the morpholino oligomer, wherein:
the CPP is of a formula selected from the group consisting of:

wherein independently for each instance R a is H or acetyl, and the CPP is conjugated to the morpholino oligomer by a linker selected from:

16. A pharmaceutical composition, comprising:
a pharmaceutically acceptable carrier and an antisense compound, wherein the antisense compound comprises:
an antisense oligomer of 8-30 morpholino subunits linked together by phosphorodiamidate linkages, wherein:
each morpholino subunit is of a formula:

wherein each B is a base pairing moiety independently selected from adenine, cytosine, guanine, uracil, thymine or hypoxanthine, wherein each B taken together comprises a sequence of at least 8 contiguous bases that is complementary to: (i) the polyCUG repeats in the 3′UTR region of dystrophia myotonica protein kinase (DMPK) mRNA in Myotonic dystrophy type 1 (DM1) or (ii) the polyCCUG repeats in the first intron of zinc finger protein 9 (ZNF9) mRNA in Myotonic dystrophy type 2 (DM2); and

Assignees

• Sarepta Therapeutics Inc

Inventors

• Kole Ryszard
• Hanson Gunnar J

Classifications

• A61P21/00

  Drugs for disorders of the muscular or neuromuscular system · CPC title

• C12N15/1137 Primary

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title

• C12N2810/40

  Vectors comprising a peptide as targeting moiety, e.g. a synthetic peptide, from undefined source · CPC title

• C12N2310/3233

  Morpholino-type ring · CPC title

• C12N15/113

  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 title