Antisense composition and method for treating muscle atrophy

• US 10,006,031 B2
• Filed: 06/08/2016
• Issued: 06/26/2018
• Est. Priority Date: 02/09/2005
• Status: Active Grant

• Current Assignee: Sarepta Therapeutics, Inc.
• Original Assignee: Sarepta Therapeutics, Inc.
• Inventors: Iversen, Patrick L., Weller, Dwight D., Timmins, Alan P.
• Primary Examiner(s): ZARA, JANE J
• Application Number:
  US15/177,244
• Publication Number:
  US 20160281092A1
• Time in Patent Office: 748 Days
• Field of Search: 435 61, 435 911, 435 9131, 435455, 435 612, 514 44, 514 11, 514 12, 514 81, 514 86, 536 231, 536 245, 530327
• CPC Class Codes: A61K 31/675 having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
• CPC Class Codes: A61K 47/645 Polycationic or polyanionic oligopeptides, polypeptides or polyamino acids, e.g. polylysine, polyarginine, polyglutamic acid or peptide TAT
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: C12N 15/1136 against growth factors, growth regulators, cytokines, lymphokines or hormones
• CPC Class Codes: C12N 2310/11 Antisense
• CPC Class Codes: C12N 2310/31 of the backbone
• CPC Class Codes: C12N 2310/3145 with the nitrogen in 3' or 5'-position
• CPC Class Codes: C12N 2310/3233 Morpholino-type ring
• CPC Class Codes: C12N 2310/3513 Protein; Peptide
• CPC Class Codes: C12N 2320/30 Special therapeutic applications
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

A method and compound for treating skeletal muscle mass deficiency in a human subject are disclosed. The composition is an oligomer of morpholino subunits and phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′ exocyclic carbon of an adjacent subunit, contains between 10-40 nucleotide bases, has a base sequence effective to hybridize to an expression-sensitive region of processed or preprocessed human myostatin RNA transcript, identified, in its processed form, by SEQ ID NO: 6, and is capable of uptake by target muscle cells in the subject. In practicing the method, the compound is administered in an amount and at a dosage schedule to produce an overall reduction in the level of serum myostatin measured in the patient, and preferably to bring the myostatin level within the a range determined for normal, healthy individuals.

Claims

1. An antisense morpholino oligonucleotide of 15-40 bases, comprising a base sequence that is complementary to at least 12 contiguous bases of SEQ ID NO:

6, wherein the antisense morpholino oligonucleotide inhibits the expression of human myostatin in a cell, and wherein the antisense morpholino oligonucleotide comprises phosphorus-containing intersubunit linkages, in accordance with the structure;

2. The antisense morpholino oligonucleotide of claim 1, wherein the antisense morpholino oligonucleotide is conjugated to an arginine-rich peptide.

3. The antisense morpholino oligonucleotide of claim 1, wherein at least 2 and no more than half of the total number of phosphorus-containing intersubunit linkages are positively charged at physiological pH.

4. The antisense morpholino oligonucleotide of claim 3, wherein

X, for any uncharged linkages, is independently alkyl, alkoxy, thioalkoxy, or an alkyl amino of the form NR₂, where each R is independently hydrogen or methyl, and X, for any positively charged linkages, is 1-piperazine.

5. The antisense morpholino oligonucleotide of claim 2, wherein the arginine rich peptide is selected from SEQ ID NOS:

7-9.

6. The antisense morpholino oligonucleotide of claim 2, wherein the arginine-rich peptide is covalently coupled at its C terminus to the 5′

end of the antisense morpholino oligonucleotide.

7. The antisense morpholino oligonucleotide of claim 1, wherein the base sequence is complementary to at least 12 contiguous bases of SEQ ID NO:

10.

8. The antisense morpholino oligonucleotide of claim 7, wherein the base sequence is SEQ ID NO:

1.

9. The antisense morpholino oligonucleotide of claim 1, wherein the base sequence is complementary to at least 12 contiguous bases of a splice site in a preprocessed human myostatin transcript.

10. The antisense morpholino oligonucleotide of claim 9, wherein the splice site in the preprocessed myostatin transcript has a sequence selected from SEQ ID NOS:

11-14.

11. The antisense morpholino oligonucleotide of claim 10, wherein the base sequence is selected from SEQ ID NOS:

2-5.