Details for Patent: 8,785,410

Title:	Antisense composition and method for treating muscle atrophy
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.
Inventor(s):	Iversen; Patrick L. (Corvallis, OR), Weller; Dwight D. (Corvallis, OR), Timmins; Alan P. (Sherwood, OR)
Assignee:	Sarepta Therapeutics, Inc. (Corvallis, OR)
Filing Date:	Jan 03, 2011
Application Number:	12/983,798
Claims:	1. A method of treating skeletal muscle mass deficiency in a human subject comprising (a) measuring blood or tissue levels of myostatin in the subject, (b) administering to the subject, a myostatin-expression-inhibiting amount of 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 preprocessed human myostatin RNA transcript, identified by SEQ ID NO:13, wherein the antisense compound includes a base sequence as set forth in SEQ ID NO:3 and is capable of effectively blocking production of a full-length processed myostatin transcript from a preprocessed myostatin transcript, (c) by said administering, forming within target muscle cells in the subject, 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.degree. C., thereby inhibiting expression of myostatin in said cells, (d) at a selected time following said administering, measuring a blood or tissue level of myostatin in the subject, and (e) repeating said administering, using the myostatin levels measured in (d) to adjust the dose or dosing schedule of the amount of antisense compound administered, if necessary, so as to reduce measured levels of myostatin over those initially measured and to maintain such levels of myostatin measured in step (d) within a range determined for normal, healthy individuals. 2. The method of claim 1, wherein the myostatin value measured in step (a) is above a selected threshold for normal healthy people. 3. The method of claim 1, wherein the morpholino subunits in the compound administered are joined by phosphorodiamidate linkages, in accordance with the structure: ##STR00002## where Y.sub.1.dbd.O, Z.dbd.O, Pj is a purine or pyrimidine base-pairing moiety effective to bind, by base-specific hydrogen bonding, to a base in a polynucleotide, and X is alkyl, alkoxy, thioalkoxy, amino or alkyl amino, including dialkylamino. 4. The method of claim 1, wherein the compound is administered as a conjugate of the compound and an arginine-rich peptide effective to promote uptake of the compound into muscle target cells. 5. The method of claim 2, in which at least 2 and no more than half of the total number of intersubunit linkages are positively charged at physiological pH. 6. The method of claim 5, wherein said morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure: ##STR00003## where Y.sub.1.dbd.O, Z.dbd.O, Pj is a purine or pyrimidine base-pairing moiety effective to bind, by base-specific hydrogen bonding, to a base in a polynucleotide, and X for the uncharged linkages is alkyl, alkoxy, thioalkoxy, or an alkyl amino of the form wherein NR.sub.2, where each R is independently hydrogen or methyl, and for the positively charged linkages, X is 1-piperazine. 7. The method of claim 4, wherein the arginine-rich peptide has one of the sequences identified as SEQ ID NOS:7-9, and the arginine-rich peptide is covalently coupled at its C terminus to the 5' end of the antisense compound. 8. The method of claim 1, wherein said administering and measuring is carried out for a selected period of at least 2 weeks. 9. The method of claim 1, wherein said administering is by oral administration. 10. An antisense composition for use in increasing skeletal muscle mass in a human subject, 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 preprocessed human myostatin transcript identified by SEQ ID NO:13, wherein the antisense compound includes a base sequence as set forth in SEQ ID NO:3 and is capable of effectively blocking production of a full-length processed myostatin transcript from a preprocessed myostatin transcript. 11. The composition of claim 10, wherein the morpholino subunits in the compound are joined by phosphorodiamidate linkages, in accordance with the structure: ##STR00004## where Y.sub.1,.dbd.O, Z.dbd.O, Pj is a purine or pyrimidine base-pairing moiety effective to bind, by base-specific hydrogen bonding, to a base in a polynucleotide, and X is alkyl, alkoxy, thioalkoxy, amino or alkyl amino, including dialkylamino. 12. The composition of claim 10, which includes said compound conjugated to an arginine-rich peptide effective to promote uptake of the compound into target muscle cells. 13. The composition of claim 10, in which at least 2 and no more than half of the total number of intersubunit linkages are positively charged at physiological pH. 14. The composition of claim 13, wherein said morpholino subunits are joined by phosphorodiamidate linkages, in accordance with the structure: ##STR00005## where Y.sub.1.dbd.O, Z.dbd.O, Pj is a purine or pyrimidine base-pairing moiety effective to bind, by base-specific hydrogen bonding, to a base in a polynucleotide, and X for the uncharged linkages is alkyl, alkoxy, thioalkoxy, or an alkyl amino of the form wherein NR.sub.2, where each R is independently hydrogen or methyl, and for the positively charged linkages, X is 1-piperazine. 15. The composition of claim 12, wherein the arginine-rich peptide has one of the sequences identified as SEQ ID NOS:7-9. 16. The composition of claim 12, arginine-rich peptide is covalently coupled at its C terminus to the 5' end of the antisense compound.