Peptide-linked morpholino antisense oligonucleotides for treatment of myotonic dystrophy

Patent metadata

Field	Value
Publication number	US-10111962-B2
Application number	US-201314431115-A
Country	US
Kind code	B2
Filing date	Sep 24, 2013
Priority date	Sep 25, 2012
Publication date	Oct 30, 2018
Grant date	Oct 30, 2018

Abstract

Provided herein are peptide-linked morpholino (PPMO) antisense oligonucleotides that target the poly CUG repeat tract in the 3′ untranslated region of the gene encoding dystrophia myotonica-protein kinase (DMPK) and methods for systemic administration of the same for the treatment of mytonic dystrophy type I (DM1).

First claim

What is claimed is:

1. A method for treating or preventing myotonic dystrophy type 1 (DM1) in an individual in need thereof comprising: systemically administering to the individual a therapeutically effective amount of a cationic peptide-linked morpholino antisense oligonucleotide comprising: (a) a morpholino antisense oligonucleotide sequence complementary to at least 3 polyCUG repeat sequences in a 3′ untranslated region (UTR) of a dystrophia myotonica protein kinase (DMPK) RNA transcript target; and (b) a spacer moiety linking the morpholino antisense oligonucleotide and a cationic peptide, the space moiety comprising

wherein administration of the cationic peptide-linked morpholino antisense oligonucleotide relieves at least one symptom of DM1 in at least two muscles.

2. The method of claim 1 , wherein the cationic peptide is 8-30 amino acid residues in length and comprises one or more subsequences selected from the group consisting of RXR, RX, RB, and RBR, wherein R is arginine, B is β-alanine, and each X is independently —NH—(CHR 1 ) n —C(O)—, wherein n is 4-6 and each R 1 is independently H or methyl such that at most two R 1 s are methyl.

3. The method of claim 1 , wherein the cationic peptide-linked morpholino antisense oligonucleotide comprises the sequence 5′-(AGC) n -3′(SEQ ID NO:3),5′-(GCA) n -3′(SEQ ID NO:4), or 5′-(CAG) n -3′(SEQ ID NO:5), wherein n is any of about 5-25.

4. The method of claim 3 , wherein the cationic peptide-linked morpholino antisense oligonucleotide further comprises 1 to 2 additional morpholino nucleotides on the 5′ and/or the 3′ end of the oligonucleotide.

5. The method of claim 3 , wherein the cationic peptide-linked morpholino antisense oligonucleotide comprises the sequence: 5′-AGCAGCAGCAGCAGCAGCAGCAGCA-3′(SEQ ID NO:6).

6. The method of claim 5 , wherein the cationic peptide-linked morpholino antisense oligonucleotide further comprises a 5′ amine modification.

7. The method of claim 1 , wherein the cationic peptide-linked morpholino antisense oligonucleotide is a phosphorodiamidate cationic peptide-linked morpholino antisense oligonucleotide.

8. The method of claim 1 , wherein the cationic peptide is separated from the morpholino antisense oligonucleotide by the spacer moiety linked at the 5′ end of the morpholino antisense oligonucleotide.

9. The method of claim 1 , wherein the muscles are skeletal muscles, smooth muscles, and/or cardiac muscle.

10. The method of claim 1 , wherein the cationic peptide-linked morpholino antisense oligonucleotide penetrates into cells of the tibialis anterior muscle, the quadriceps muscle, and/or the gastrocnemius muscle.

11. The method of claim 1 , wherein said at least one symptom of DM1 is myotonia.

12. The method of claim 1 , wherein said at least one symptom of DM1 is aggregation of musclebind-like-1 (MBNL-1) protein in ribonuclear foci within myonuclei.

13. The method of claim 1 , wherein said at least one symptom of DM1 is abnormal splicing of at least one RNA transcript in muscle cells.

14. The method of claim 13 , wherein said at least one RNA transcript is selected from the group consisting of: Serca-1, m-Titin, Zasp, and CIC-1.

15. The method of claim 1 , wherein systemic administration of the cationic peptide-linked morpholino antisense oligonucleotide is performed intravenously, intraperitoneally, or subcutaneously.

16. The method of claim 1 , wherein the cationic peptide-linked morpholino antisense oligonucleotide is administered to the individual weekly.

17. The method of claim 1 , wherein the cationic peptide-linked morpholino antisense oligonucleotide is administered to the individual for at least one week.

18. The method of claim 1 , wherein the cationic peptide-linked morpholino antisense oligonucleotide is administered with a pharmaceutically acceptable excipient.

19. The method of claim 1 , wherein the individual is a human.

20. A method for treating or preventing myotonic dystrophy type 1(DM1) in an individual in need thereof comprising: systemically administering to the individual a therapeutically effective amount of a cationic peptide-linked morpholino antisense oligonucleotide comprising: (a) morpholino antisense oligonucleotide sequence complementary to at least 3polyCUG repeat sequences in a 3′ untranslated region (UTR) of a dystrophia myotonica protein kinase (DMPK) RNA transcript target; (b) a spacer moiety linking the morpholino antisense oligonucleotide and a cationic peptide, the space moiety comprising

and
(c) the, cationic peptide comprising the amino acid sequence Ac(RXRRBR) 2 XB-(SEQ ID NO:1), wherein Ac is acetyl, R is arginine, B is β-alanine, and each X is independently —NH—(CHR 1 ) n —,C(O)—, wherein n is 4-6 and each R 1 is independently H or methyl such that at most two R 1 s are methyl, and wherein administration of the cationic peptide-linked rnorpholino antisense oligonucleotide relieves at least one symptom of DM1 in at least two muscles.

21. A method for treating or preventing myotonic dystrophy type 1(DM1) in an individual in need thereof comprising: systemically administering to the individual a therapeutically effective amount of a cationic peptide-linked morpholino antisense oligonucleotide comprising: (a) a morpholino antisense oligonucleotide sequence complementary to at least 3polyCUG repeat sequences in a 3′ untranslated region (UTR) of a dystrophia myotonica protein kinase (DMPK) RNA transcript target: (b) a spacer moiety linking the morpholino antisense oligonucleotide and a cationic peptide, the space moiety comprising

and
(c) the, cationic peptide comprising the amino acid sequence Ac(RXR) 4 XB-(SEQ ID NO:2), wherein Ac is acetyl, R is arginine, B is β-alanine, and each X is independently —NH—(CHR 1 ) n —C(O)—, wherein n is 4-6 and each R 1 is independently H or methyl such that at most two R 1 s are methyl, and wherein administration of the cationic peptide-linked morpholino antisense oligonucleotide relieves at least one symptom of DM1 in at least two muscles.

Assignees

• Genzyme Corp

Inventors

• Leger Andrew
• Wentworth Bruce
• Nelson Carol A
• Weeden Timothy E
• Clayton Nicholas
• Cheng Seng

Classifications

• A61P21/02

  Muscle relaxants, e.g. for tetanus or cramps · CPC title

• A61P21/00

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

• A61K47/64 Primary

  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 title

• C12N2320/32

  Special delivery means, e.g. tissue-specific · CPC title

• C12N2310/3513

  Protein; Peptide · CPC title