Antisense composition and method for treating muscle atrophy

• US 7,888,012 B2
• Filed: 05/11/2006
• Issued: 02/15/2011
• Est. Priority Date: 02/09/2005
• Status: Active Grant

• Current Assignee: Avi Biopharma, Inc.
• Original Assignee: Avi Biopharma, Inc.
• Inventors: Iversen, Patrick L., Weller, Dwight D., Timmins, Alan P.
• Primary Examiner(s): ZARA, JANE J
• Application Number:
  US11/433,724
• Publication Number:
  US 20070122821A1
• Time in Patent Office: 1,741 Days
• Field of Search: 435/6, 435/91.1, 435/91.31, 435/455, 514/44, 536/23.1, 536/24.5
• US Class Current: 435/6.16
• 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 composition, comprising a substantially uncharged antisense compound

(i) composed of morpholino subunits and phosphorus-containing intersubunit linkages joining a morpholino nitrogen of one subunit to a 5′

exocyclic carbon of an adjacent subunit,

(ii) capable of uptake by target muscle cells in the subject,

(iii) containing between 10-40 nucleotide bases, and

(iv) having a base sequence that is complementary to at least 12 contiguous bases in a target region of processed human myostatin transcript identified by SEQ ID NO;

10, where hybridizing of the compound to the target region is effective to block translation of processed human myostatin RNA transcript, and

conjugated to the antisense compound, an arginine-rich peptide has the sequence identified by SEQ ID NO;

9.

2. The composition of claim 1, wherein the morpholino subunits in the compound are joined by phosphorodiamidate linkages, in accordance with the structure:

3. The composition of claim 1, in which a least 2 and no more than half of the total number of intersubunit linkages are positively charged at physiological pH.

4. The composition of claim 3, wherein said morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure:

5. The composition of claim 1, wherein the arginine-rich peptide is covalently coupled at its C terminus to the 5′

end of the antisense compound.

6. The composition of claim 1, wherein the antisense compound includes a base sequence identified by the sequence SEQ ID NO:

1.

7. A method of measuring myostatin expression levels in a mammalian subject, comprising

(a) administering to the subject, a myostatin-expression-inhibiting amount of the substantially uncharged antisense compound of claim 1, which is capable of forming with a such myostatin RNA transcript, a base-paired heteroduplex structure composed of human myostatin RNA transcript and the antisense compound and having a Tm of dissociation of at least 45°

C.,

(b) within about 8-72 hours following said administering, analyzing a body-fluid sample obtained from the subject for the presence of a heteroduplex composed of said antisense compound and a complementary region of said myostatin RNA transcript, to determine the concentration of transcript in said sample.