Oligonucleotide analogues targeting human LMNA

1. An antisense oligonucleotide for use in modulating aberrant splicing of a human LMNA pre-mRNA, the oligonucleotide being 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, and:

(i) having a substantially uncharged, nuclease resistant backbone;

(ii) capable of uptake by mammalian host cells;

(iii) containing between about 12-40 nucleotide bases; and

(iv) having a targeting sequence of at least about 12 contiguous subunits complementary to exon 10, intron 10, exon 11, or combinations thereof, of a human LMNA pre-mRNA; and

wherein the targeting sequence comprises any one of SEQ ID NOs: 3-7, 11, or 14-16.

2. The oligonucleotide of claim 1 , where the targeting sequence consists essentially of SEQ ID NO: 4.

3. The oligonucleotide of claim 1 , where the targeting sequence consists essentially of SEQ ID NO: 11.

4. The oligonucleotide of claim 1 , where the morpholino subunits in the oligonucleotide are joined by phosphorodiamidate linkages, in accordance with the following structure:

wherein Z is S or O,

X═NR 1 R 2 or OR 6 ,

Y═O or NR 7 ,

and each said linkage is selected from:

(a) uncharged linkage (a), wherein each of R 1 , R 2 , R 6 , and R 7 is independently selected from hydrogen and lower alkyl;

(b1) cationic linkage (b1), wherein X═NR 1 R 2 and Y═O, and NR 1 R 2 represents an optional substituted piperazino group, such that R 1 R 2 ═

—CHRCHRN(R 3 )(R 4 )CHRCHR—, wherein

each R 4 is H, CH 3 or null, and

R3 is selected from H, lower alkyl, C(═NH)NH 2 , Z—L—NHC(═NH)NH 2 , and [C(O)CHR′NH] m H, wherein where Z is carbonyl (C(O)) or a direct bond, L is an optional linker up to 18 atoms in length having bonds selected from alkyl, alkoxy, and alkylamino, R′ is a side chain of a naturally occurring amino acid or a one- or two-carbon homolog thereof, and m is 1 to 6;

(b2) cationic linkage (b 2 ), wherein X═NR 1 R 2 and Y═O, R 1 ═H or CH 3 , and R 2 ═LNR 3 R 4 R 5 , wherein L, R 3 , and R 4 are defined as above, and R 5 is H, lower alkyl, or lower (alkoxy)alkyl; and

(b3) cationic linkage (b 3 ), wherein Y═NR 7 and X═OR 6 , and R7═LNR 3 R 4 R 5 . wherein L, R 3 , and R 4 and R 5 are defined as above, and R 6 is H or lower alkyl; and at least one said linkage is selected from cationic linkages (b1), (b2), and (b3).

5. The oligonucleotide of claim 4 , where each of R 1 and R 2 , in linkages of type (a), is methyl.

6. The oligonucleotide of claim 4 , where at least one linkage is of type (b 1 ), where each R is H, R 4 is H, CH 3 , or an electron pair, and R 3 is selected from H, CH 3 , C(═NH)NH 2 , and C(O)—L—NHC(═NH)NH 2 .

7. The oligonucleotide of claim 1 , where the antisense oligonucleotide is covalently attached to a cell-penetrating peptide.

8. The oligonucleotide of claim 7 , where the cell-penetrating peptide is an arginine-rich peptide.

9. The oligonucleotide of claim 8 , where the peptide is attached at its C-terminus to the 3′ end of the oligonucleotide through a one- or two-amino acid linker.

10. An antisense oligonucleotide, or a pharmaceutical composition comprising the same, wherein the oligonucleotide modulates aberrant splicing of a human LMNA pre-mRNA, the oligonucleotide comprising a backbone, the backbone comprising a sequence of morpholino ring structures joined by intersubunit linkages, the intersubunit linkages joining a 3′-end of one morpholino ring structure to a 5′-end of an adjacent morpholino ring structure, wherein each morpholino ring structure is bound to a base-pairing moiety, such that the oligonucleotide can bind in a sequence-specific manner to a target nucleic acid, comprising a targeting sequence that is complementary to at least 12 bases of a sequence set forth in any one of SEQ ID NO:1-34, or which comprises any one or more of SEQ ID NOS: 3-34, wherein the intersubunit linkages have the following general structure (I):

or a salt or isomer thereof, and wherein each of the intersubunit linkages (I) are independently linkage (A) or linkage (B):

wherein for linkage (A):

W is, at each occurrence, independently S or O;

X is, at each occurrence, independently —N(CH 3 ) 2 , —NR 1 R 2 , —OR 3 or ;

Y is, at each occurrence, independently O or —NR 2 ,

R 1 is, at each occurrence, independently hydrogen or methyl;

R 2 is, at each occurrence, independently hydrogen or —LNR 4 R 5 R 7 ;

R 3 is, at each occurrence, independently hydrogen or C 1 -C 6 alkyl;

R 4 is, at each occurrence, independently hydrogen, methyl, —C(═NH)NH 2 , —Z—L—NHC(═NH)NH 2 or —[C(O)CHR′NH] m H, where Z is carbonyl (C(O)) or a direct bond, R′ is a side chain of a naturally occurring amino acid or a one- or two-carbon homolog thereof, and m is 1 to 6;

R 5 is, at each occurrence, independently hydrogen, methyl or an electron pair;

R 6 is, at each occurrence, independently hydrogen or methyl;

R 7 is, at each occurrence, independently hydrogen C 1 -C 6 alkyl or C 1 -C 6 alkoxyalkyl;

L is an optional linker up to 18 atoms in length comprising alkyl, alkoxy or alkylamino groups, or combinations thereof; and

wherein for linkage (B):

W is, at each occurrence, independently S or O;

X is, at each occurrence, independently —NR 8 R 9 or —OR 3 ; and

Y is, at each occurrence, independently O or —NR 10 ,

R 8 is, at each occurrence, independently hydrogen or C 2 -C 12 alkyl;

R 9 is, at each occurrence, independently hydrogen, C 1 -C 12 alkyl, C 1 -C 12 aralkyl or aryl;

R 10 is, at each occurrence, independently hydrogen, C 1 -C 12 alkyl or —LNR 4 R 5 R 7 ;

wherein R 8 and R 9 may join to form a 5-18 membered mono or bicyclic heterocycle or R 8 , R 9 or R 3 may join with R 10 to form a 5-7 membered heterocycle, and wherein when X is 4-piparazino, X has the following structure (III):

wherein:

R 11 is, at each occurrence, independently C 2 -C 12 alkyl, C 1 -C 12 aminoalkyl, C 1 -C 12 alkylcarbonyl, aryl, heteroaryl or heterocyclyl; and

R is, at each occurrence, independently an electron pair, hydrogen or C 1 -C 12 alkyl; and

R 12 is, at each occurrence, independently, hydrogen, C 1 -C 12 alkyl, C 1 -C 12 aminoalkyl, —NH 2 , —NR 13 R 14 , —NR 13 R 14 R 15 , C 1 -C 12 alkylcarbonyl, oxo, —CN, trifluoromethyl, amidyl, amidinyl, amidinylalkyl, amidinylalkylcarbonyl guanidinyl, guanidinylalkyl, guanidinylalkylcarbonyl, cholate, deoxycholate, aryl, heteroaryl, heterocycle, —SR 13 or C 1 -C 12 alkoxy, wherein R 13 , R 14 and R 15 are, at each occurrence, independently C 1 -C 12 alkyl;

wherein at least one of the intersubunit linkages is linkage (B); and

wherein the targeting sequence comprises any one of SEQ ID NOs: 3-7, 11, or 14-16.

11. The oligonucleotide of claim 10 , where the targeting sequence consists essentially of SEQ ID NO: 4.

12. The oligonucleotide of claim 10 , where the targeting sequence consists essentially of SEQ ID NO: 11.