PHOSPHORODIAMIDATE MORPHOLINO OLIGOMER CONJUGATES
Novel antisense oligonucleotide conjugates that cause skipping of an exon in the human dystrophin gene and their use in a method of treating muscular dystrophy in a patient suffering from Duchenne muscular dystrophy (DMD) are described.
1 . An antisense oligomer conjugate of Formula (I):
or a pharmaceutically acceptable salt thereof,
wherein:
n is 1-40;
each Nu is a nucleobase, which, taken together, form a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA;
T′ is a moiety selected from:
wherein
R 100 is selected from the group consisting of RRRRRG-, RRRRG-, RRRG-, RRG-, RG-, and G-, wherein R is arginine and G is glycine,
R 200 is hydrogen, and
R 1 is C 1 -C 6 alkyl.
2 . The antisense oligomer conjugate of claim 1 , or a pharmaceutically acceptable salt thereof, wherein the targeting sequence is complementary to an exon 51 annealing site in the dystrophin pre-mRNA designated as H51A(+66+95).
3 . The antisense oligomer conjugate of claim 1 , or a pharmaceutically acceptable salt thereof, wherein the targeting sequence is complementary to an exon 45 annealing site in the dystrophin pre-mRNA designated as H45A(−03+19).
4 . The antisense oligomer conjugate of claim 1 , or a pharmaceutically acceptable salt thereof, wherein the targeting sequence is complementary to an exon 53 annealing site in the dystrophin pre-mRNA designated as H53A(+36+60).
5 . The antisense oligomer conjugate of any one of claims 1-4 , or a pharmaceutically acceptable salt thereof, wherein each Nu is independently selected from cytosine (C), guanine (G), thymine (T), adenine (A), 5-methylcytosine (5mC), uracil (U), and hypoxanthine (I).
6 . The antisense oligomer conjugate of any one of claims 1-5 , or a pharmaceutically acceptable salt thereof, wherein T′ is a moiety:
wherein R 200 is hydrogen.
7 . The antisense oligomer conjugate of any one of claims 1-6 , or a pharmaceutically acceptable salt thereof, wherein R 100 is RRRRRG-.
8 . The antisense oligomer conjugate of any one of claims 1-6 , or a pharmaceutically acceptable salt thereof, wherein R 100 is RRRRG-.
9 . The antisense oligomer conjugate of any one of claims 1-6 , or a pharmaceutically acceptable salt thereof, wherein R 100 is RRRG-.
10 . The antisense oligomer conjugate of any one of claims 1-6 , or a pharmaceutically acceptable salt thereof, wherein R 100 is RRG-.
11 . The antisense oligomer conjugate of any one of claims 1-6 , or a pharmaceutically acceptable salt thereof, wherein R 100 is RG-.
12 . The antisense oligomer conjugate of any one of claims 1-6 , or a pharmaceutically acceptable salt thereof, wherein R 100 is G-.
13 . The antisense oligomer conjugate of any one of claims 1-12 , having the Formula (V):
or a pharmaceutically acceptable salt thereof, wherein
each Nu is a nucleobase, which, taken together, form a targeting sequence that is complementary to an exon annealing site in the dystrophin pre-mRNA, and
m is 0, 1, 2, 3, 4, or 5.
14 . The antisense oligomer conjugate of any one of claims 1-13 , having the Formula (VA):
or a pharmaceutically acceptable salt thereof, wherein m is 0, 1, 2, 3, 4, or 5, and
each Nu from 1 to 30 and 5′ to 3′ is:
Position
No. 5′
to 3′
Nu
1
C
2
T
3
C
4
C
5
A
6
A
7
C
8
A
9
T
10
C
11
A
12
A
13
G
14
G
15
A
16
A
17
G
18
A
19
T
20
G
21
G
22
C
23
A
24
T
25
T
26
T
27
C
28
T
29
A
30
G
wherein A is
C is
G is
and T is
15 . The antisense oligomer conjugate of any one of claims 1-12 , having the Formula (VII):
or a pharmaceutically acceptable salt thereof, wherein
each Nu is a nucleobase, which, taken together, form a targeting sequence that is complementary to an exon annealing site in the dystrophin pre-mRNA, and
m is 0, 1, 2, 3, 4, or 5.
16 . The antisense oligomer conjugate of any one of claims 1-12 or 15 , having the Formula (VIIA):
or a pharmaceutically acceptable salt thereof, wherein m is 0, 1, 2, 3, 4, or 5, and
each Nu from 1 to 22 and 5′ to 3′ is:
Position
No. 5′
to 3′
Nu
1
C
2
A
3
A
4
T
5
G
6
C
7
C
8
A
9
T
10
C
11
C
12
T
13
G
14
G
15
A
16
G
17
T
18
T
19
C
20
C
21
T
22
G
wherein A is
C is
G is
and T is
17 . The antisense oligomer conjugate of any one of claims 1-12 , having the Formula (IX):
or a pharmaceutically acceptable salt thereof, wherein
each Nu is a nucleobase, which, taken together, form a targeting sequence that is complementary to an exon annealing site in the dystrophin pre-mRNA, and
m is 0, 1, 2, 3, 4, or 5.
18 . The antisense oligomer conjugate of any one of claims 1-12 or 17 , having the Formula (IXA):
or a pharmaceutically acceptable salt thereof, wherein m is 0, 1, 2, 3, 4, or 5, and
each Nu from 1 to 25 and 5′ to 3′ is:
Position
No. 5′
to 3′
Nu
1
G
2
T
3
T
4
G
5
C
6
C
7
T
8
C
9
C
10
G
11
G
12
T
13
T
14
C
15
T
16
G
17
A
18
A
19
G
20
G
21
T
22
G
23
T
24
T
25
C
wherein A is
C is
G is
and T is
19 . The antisense oligomer conjugate of any one of claims 13-18 , or a pharmaceutically acceptable salt thereof, wherein m is 0.
20 . The antisense oligomer conjugate of any one of claims 13-18 , or a pharmaceutically acceptable salt thereof, wherein m is 1.
21 . The antisense oligomer conjugate of any one of claims 13-18 , or a pharmaceutically acceptable salt thereof, wherein m is 2.
22 . The antisense oligomer conjugate of any one of claims 13-18 , or a pharmaceutically acceptable salt thereof, wherein m is 3.
23 . The antisense oligomer conjugate of any one of claims 13-18 , or a pharmaceutically acceptable salt thereof, wherein m is 4.
24 . The antisense oligomer conjugate of any one of claims 13-18 , or a pharmaceutically acceptable salt thereof, wherein m is 5.
25 . The antisense oligomer conjugate of any one of claims 1-24 , wherein the antisense oligomer conjugate is a free base.
26 . The antisense oligomer conjugate of any one of claims 1-24 , wherein the antisense oligomer conjugate is a pharmaceutically acceptable salt.
27 . The antisense oligomer conjugate of any one of claims 1-24 or 26 , wherein the antisense oligomer conjugate is a hydrochloride salt.
28 . A pharmaceutical composition, comprising an antisense oligonucleotide conjugate of any one of claims 1-27 , or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.
29 . The pharmaceutical composition of claim 28 , wherein the pharmaceutical composition is formulated for parenteral use.
30 . A method of treating a patient with Duchenne muscular dystrophy (DMD) in need thereof who has a mutation that is amenable to exon skipping, comprising administering to the patient an antisense oligomer conjugate of any one of claims 1-27 , or a pharmaceutically acceptable salt thereof.
31 . The method of claim 30 , wherein the antisense oligomer conjugate causes skipping of an exon in the human dystrophin gene.
32 . The method of claim 30 or 31 , wherein the exon is chosen from exon 44, 45, 50, 51, 52, or 53.
33 . The method of any one of claims 30-32 , wherein the exon is chosen from exon 45, 51, or 53.
34 . A method of treating a patient with Duchenne muscular dystrophy (DMD) with an antisense oligomer conjugate, comprising administering to the patient an antisense oligomer conjugate of any one of claims 1-27 , or a pharmaceutically acceptable salt thereof.
35 . A method of treating a patient with Duchenne muscular dystrophy (DMD) in need thereof who has a mutation that is amenable to exon skipping, comprising administering to the patient a composition of claim 28 or 29 , and a pharmaceutically acceptable carrier.
36 . The method of claim 35 , wherein the antisense oligomer conjugate causes skipping of an exon in the human dystrophin gene.
37 . The method of claim 35 or 36 , wherein the exon is chosen from exon 44, 45, 50, 51, 52, or 53.
38 . The method of any one of claims 35-37 , wherein the exon is chosen from exon 45, 51, or 53.
39 . A method of treating a patient with Duchenne muscular dystrophy (DMD) with an antisense oligomer conjugate, comprising administering to the patient a composition of claims 28 or 29 , and a pharmaceutically acceptable carrier.
40 . A method of treating a patient with DMD in need thereof who has a mutation that is amenable to exon 51 skipping, comprising administering to the patient an antisense oligomer conjugate having Formula (VI):
or a pharmaceutically acceptable salt thereof, wherein m is 0, 1, 2, 3, 4, or 5.
41 . A method of treating a patient with DMD in need thereof who has a mutation that is amenable to exon 45 skipping, comprising administering to the patient an antisense oligomer conjugate having Formula (VIII):
or a pharmaceutically acceptable salt thereof, wherein m is 0, 1, 2, 3, 4, or 5.
42 . A method of treating a patient with DMID in need thereof who has a mutation that is amenable to exon 53 skipping, comprising administering to the patient an antisense oligomer conjugate having Formula (X):
or a pharmaceutically acceptable salt thereof, wherein m is 0, 1, 2, 3, 4, or 5.
43 . An antisense oligomer conjugate according to any one of claims 1-27 , or a pharmaceutically acceptable salt thereof, for use in the treatment of Duchenne muscular dystrophy (DMD) in a patient who has a mutation that is amenable to exon skipping.
44 . The antisense oligomer conjugate for use according to claim 43 , wherein the antisense oligomer conjugate causes skipping of an exon in the human dystrophin gene.
45 . The antisense oligomer conjugate for use according to claim 43 or 44 , wherein the exon is chosen from exon 44, 45, 50, 51, 52, or 53.
46 . The antisense oligomer conjugate for use according to any one of claims 43-45 , wherein the exon is chosen from exon 45, 51, or 53.
47 . A pharmaceutical composition comprising an antisense oligonucleotide conjugate according to any one of claims 1-27 , or a pharmaceutically acceptable salt thereof, for use in the treatment of Duchenne muscular dystrophy (DMD) in a patient who has a mutation that is amenable to exon skipping.
48 . The pharmaceutical composition for use according to claim 47 , wherein the antisense oligomer conjugate causes skipping of an exon in the human dystrophin gene.
49 . The pharmaceutical composition for use according to claim 47 or 48 , wherein the exon is chosen from exon 44, 45, 50, 51, 52, or 53.
50 . The pharmaceutical composition for use according to any one of claims 47-49 , wherein the exon is chosen from exon 45, 51, or 53.
51 . The pharmaceutical composition for use according to any one of claims 47-50 , wherein the pharmaceutical composition is formulated for parenteral use.
52 . An antisense oligomer conjugate according to any one of claims 1-27 , or a pharmaceutically acceptable salt thereof, for use as a medicament.