Antisense-induced exon exclusion in myostatin

1. An antisense oligomer compound of consisting of a sequence selected from SEQ ID NOS: 2-5, 10-15, and 17-30, wherein X is independently selected from uracil (U) or thymine (T), comprising:

at least one subunit that is a nucleotide analog having (i) a modified internucleoside linkage, (ii) a modified sugar moiety, or (iii) a combination of the foregoing.

2. The antisense oligomer compound of claim 1 , wherein the modified internucleoside linkage is selected from a phosphorothioate internucleoside linkage, a phosphoramidate internucleoside linkage, a phosphorodiamidate internucleoside linkage, or a phosphorodiamidate wherein the phosphorous atom is covalently bonded to a (1,4-piperazin)-1-yl moiety, a substituted (1,4-piperazin)-1-yl moiety, a 4-aminopiperidin-1-yl moiety, or a substituted 4-aminopiperidin-1-yl moiety.

3. The antisense oligomer compound of claim 1 , further comprising an arginine-rich cell-penetrating peptide covalently bonded to the 3′ or the 5′ end of the antisense oligomer compound.

4. The antisense oligomer compound of claim 1 , wherein a nucleobase of each of the subunits is independently adenine, guanine, thymine, uracil, cytosine, inosine, hypoxanthine, 2,6-diaminopurine, 5-methyl cytosine, C5-propynyl-modified pyrimidines, or 10-(9-(aminoethoxy)phenoxazinyl).

5. A compound of formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

each Nu is a nucleobase which taken together form a targeting sequence;

Z is an integer from 15 to 25;

each Y is independently selected from O and —NR 4 , wherein each R 4 is independently selected from H, C 1 -C 6 alkyl, aralkyl, —C(═NH)NH 2 , —C(O)(CH 2 ) n NR 5 C(═NH)NH 2 ,

—C(O)(CH 2 ) 2 NH 5 C(O)(CH 2 )NR 5 C(═NH)NH 2 , and G, wherein R 5 is selected from H and C 1 -C 6 alkyl and n is an integer from 1 to 5;

T is selected from OH and a moiety of the formula:

wherein:

A is selected from —OH, —N(R 7 ) 2 , and R 1 wherein:

each R 7 is independently selected from H and C 1 -C 6 alkyl, and

R 6 is selected from OH, —N(R 9 )CH 2 C(O)NH 2 , and a moiety of the formula:

wherein:

R 9 is selected from H and C 1 -C 6 alkyl; and

R 10 is selected from G, —C(O)—R 11 OH, acyl, trityl, 4-methoxytrityl,

—C(═NH)NH 2 , —C(O)(CH 2 ) m NR 12 C(═NH)NH 2 , and

—C(O)(CH 2 ) 2 NHC(O)(CH 2 ) 5 NR 12 C(═NH)NH 2 , wherein:

m is an integer from 1 to 5,

R 11 is of the formula —(O-alkyl) y — wherein y is an integer from 3 to 10 and each of the y alkyl groups is independently selected from C 2 -C 6 alkyl; and

R 12 is selected from H and C 1 -C 6 alkyl;

each instance of R 1 is independently selected from:

—N(R 13 ) 2 , wherein each R 13 is independently selected from H and C 1 -C 6 alkyl;

a moiety of formula (II):

wherein:

R 15 is selected from H, G, C 1 -C 6 alkyl, —C(═NH)NH 2 ,

—C(O)(CH 2 ) q NR 18 C(═NH)NH 2 , and

—C(O)(CH 2 ) 2 NHC(O)(CH 2 ) 5 NR 18 C(═NH)NH 2 , wherein:

R 18 is selected from H and C 1 -C 6 alkyl; and

q is an integer from 1 to 5; and

each R 17 is independently selected from H and methyl; and

a moiety of formula(III):

wherein:

R 19 is selected from H, C 1 -C 6 alkyl, —C(═NH)NH 2 , —C(O)(CH 2 ) r NR 22 C(═NH)NH 2 ,

—C(O)CH(NH 2 )(CH 2 ) 3 NHC(═NH)NH 2 ,

—C(O)(CH 2 ) 2 NHC(O)(CH 2 ) 5 NR 22 C(═NH)NH 2 , —C(O)CH(NH 2 )(CH 2 ) 4 NH 2 and G,

wherein:

R 22 is selected from H and C 1 -C 6 alkyl; and

r is an integer from 1 to 5,

R 20 is selected from H and C 1 -C 6 alkyl; and

R 2 is selected from H, G, acyl, trityl, 4-methoxytrityl, C 1 -C 6 alkyl, —C(═NH)NH 2 , —C(═NH)NH 2 , —C(O)—R 23 ,

—C(O)(CH 2 ) s NR 24 C(═NH)NH 2 ,

—C(O)(CH 2 ) 2 NHC(O)(CH 2 ) 5 NR 24 C(═NH)NH 2 , —C(O)CH(NH 2 )(CH 2 ) 3 NHC(═NH)NH 2 , and a moiety of the formula:

wherein,

R 23 is of the formula —(O-alkyl) v —OH wherein v is an integer from 3 to 10 and each of the v alkyl groups is independently selected from C 2 -C 6 alkyl; and

R 24 is selected from H and C 1 -C 6 alkyl;

s is an integer from 1 to 5;

L is selected from —C(O)(CH 2 ) 6 C(O)—and —C(O)(CH 2 ) 2 S 2 (CH 2 ) 2 C(O)—; and

each R 25 is of the formula —(CH 2 ) 2 OC(O)N(R 26 ) 2 wherein each R 26 is of the formula —(CH 2 ) 6 NHC(═NH)NH 2 ,

wherein G is a cell penetrating peptide (“CPP”) and linker moiety selected from —C(O)(CH 2 ) 5 NH—CPP, —C(O)(CH 2 ) 2 NH—CPP, —C(O)(CH 2 ) 2 NHC(O)(CH 2 ) 5 NH—CPP, and —C(O)CH 2 NH—CPP, or G is of the formula:

wherein the CPP is attached to the linker moiety by an amide bond at the CPP carboxy terminus, with the proviso that up to one instance of G is present, and

wherein the targeting sequence consists of a sequence selected from SEQ ID NOS: 2-5, 10-15, and 17-30 and wherein X is independently selected from uracil (U) or thymine (T).

6. The compound of claim 5 , wherein each Nu is independently adenine, guanine, thymine, uracil, cytosine, inosine, hypoxanthine, 2,6-diaminopurine, 5-methyl cytosine, C5-propynyl-modified pyrimidines, or 10-(9-(aminoethoxy)phenoxazinyl).

7. The compound of claim 5 , wherein the targeting sequence is selected from SEQ ID NOS: 24 or 29.

8. The compound of claim 5 , wherein Y is O, R 2 is selected from H or G, R 3 is selected from an electron pair or H.

9. The compound of claim 8 , wherein R 2 is G, and wherein the CPP is of a sequence selected from SEQ ID NOS: 32-47.

10. The compound of claim 5 , wherein each R 1 is —N(CH 3 ) 2 .

11. The compound of claim 5 , wherein T is of the formula:

12. The compound of claim 5 , wherein T is of the formula:

wherein A is —N(CH 3 ) 2 , and R 6 is of the formula:

wherein R 10 is G, and

wherein G is —C(O)CH 2 NH—CPP.

13. The compound of claim 12 , wherein the CPP is of a sequence selected from SEQ ID NOS: 32-47.

14. The antisense oligomer compound of claim 1 , wherein the targeting sequence is selected from SEQ ID NOS:17-29.