COMPOSITIONS TARGETING SODIUM CHANNEL 1.6
The invention provides therapeutic compositions that include an antisense oligonucleotide (ASO) complementary to an identified target on a Nav channel mRNA. The ASO hybridizes to its target RNA and forms a duplex that recruits RNase H to degrade the RNA, thereby downregulating Nav channel synthesis, which inhibits the neuron's ability to contribute to certain conditions such as epilepsy and pain perception. The ASO binds to one of the specific identified targets, and may be provided as a gapmer that includes a central DNA segment flanked by modified RNA wings.
1 . A composition comprising:
an oligonucleotide that hybridizes to an RNA encoding a sodium channel protein along a segment of the RNA that is at least about 75% complementary to one of SEQ ID NOs: 1-156 to thereby prevent translation of the RNA into the sodium channel protein.
2 . The composition of claim 1 , wherein the oligonucleotide hybridizes to, and knocks down expression of, Nav1.6 pre-mRNA or mRNA.
3 . The composition of claim 1 , wherein a sequence of bases in the oligonucleotide has at least about 80% identity to one of SEQ ID NOs: 1-156.
4 . The composition of claim 1 , wherein a sequence of bases in the oligonucleotide is at least about 95% identical to one of SEQ ID NOs: 1-156, wherein the oligonucleotide can hybridize to, and induce RNase cleavage of, Nav1.6 pre-mRNA or mRNA.
5 . The composition of claim 1 , wherein the composition comprises a plurality of therapeutic oligonucleotides each having a base sequence at least about 80% identical to one of SEQ ID NOs: 1-156, wherein each of the therapeutic oligonucleotides has a gapmer structure comprising a central DNA segment flanked by modified RNA wings, wherein the plurality of therapeutic oligonucleotides are provided in a solution or carrier formulated for intrathecal injection.
6 . The composition of claim 1 , wherein the oligonucleotide comprises two wings flanking a central region of at least about 9 DNA bases.
7 . The composition of claim 1 , wherein at least one end of the oligonucleotide comprises modified RNA bases.
8 . The composition of claim 7 wherein each modified RNA base is selected from the group consisting of 2′-O-methoxyethyl RNA and 2′-O-methyl RNA.
9 . The composition of claim 1 , wherein the oligonucleotide comprises at least about 15 bases.
10 . The composition of claim 1 , wherein the oligonucleotide comprises between about 15 and about 25 bases.
11 . The composition of claim 1 , wherein the oligonucleotide has a backbone comprising a plurality of phosphorothioate bonds.
12 . The composition of claim 1 , wherein the oligonucleotide has a base sequence that has been screened and determined to not meet a threshold match for any non-target transcripts in humans.
13 . The composition of claim 1 , wherein the composition comprises a plurality of copies of a therapeutic oligonucleotide having a base sequence at least about 95% identical to one of SEQ ID NOs: 1-156, wherein the therapeutic oligonucleotide has a gapmer structure comprising a central twelve-base DNA segment flanked by two wings of four 2′-O-methoxyethyl RNA bases and a backbone of phosphorothioate linkages.
14 . The composition of claim 1 , wherein when the composition is delivered to cells in vitro, the cells exhibit a dose-dependent knockdown of Nav1.6.
15 . The composition of claim 1 , wherein the oligonucleotide has a base sequence with at least about a 90% match to one of SEQ ID NO: 1-156, with bases linked only by phosphorothioate linkages, the oligonucleotide further comprising a central 12 DNA bases flanked by a 5′ wing and a 3′ wing, the 5′ wing and the 3′ wing each comprising four consecutive 2′ modified RNA bases.
16 . The composition of claim 15 , wherein the oligonucleotide has a base sequence with at least about a 90% match to one of SEQ ID NO: 16, 41, 44, 45, 100, 117, 124, 125, 126, 128, 129, 130, 133, 134, 135, 138, 139, 142, 143, and 144.
17 . The composition of claim 1 , wherein the oligonucleotide has a base sequence matching one of SEQ ID NO: 1-156, with a majority of inter-base linkages comprising phosphorothioate linkages, the oligonucleotide further comprising a central 12 DNA bases flanked by a 5′ wing and a 3′ wing, the 5′ wing and the 3′ wing each comprising four consecutive 2′-MOE RNA bases.
18 . The composition of claim 17 , wherein the oligonucleotide has a base sequence matching one of SEQ ID NO: 16, 41, 44, 45, 100, 117, 124, 125, 126, 128, 129, 130, 133, 134, 135, 138, 139, 142, 143, and 144.
19 . The composition of claim 1 , wherein the oligonucleotide hybridizes to a location within the first 3700 bases of an SCN8A transcript.
20 . The composition of claim 1 , wherein the oligonucleotide has a sequence of one selected from the group consisting of SEQ ID NO: 16; SEQ ID NO: 41; SEQ ID NO: 44; SEQ ID NO: 45; SEQ ID NO: 117; SEQ ID NO: 124; SEQ ID NO: 126; SEQ ID NO: 129; SEQ ID NO: 133; SEQ ID NO: 135; SEQ ID NO: 138; SEQ ID NO: 139; SEQ ID NO: 142; SEQ ID NO: 143; or SEQ ID NO: 144.
21 . The composition of claim 20 , further wherein the oligonucleotide is a gapmer with a 12 base DNA central segment flanked by two 2′-MOE RNA wings, in which each wing has one, two, or three phosphodiester linkages with remaining inter-base linkages being phosphorothioate, and wherein all cytosine bases have a 5-methyl modification.
22 . The composition of claim 1 , wherein the oligonucleotide knocks down expression of an SCN8A transcript to an amount about 50 to 90% compared to an un-treated control.
23 . The composition of claim 1 , further wherein the oligonucleotide is one selected from the group consisting of:
SEQ ID NO: 135 in a gapmer with a 12 base DNA central segment flanked by 2′-MOE RNA wings, in which the 2d, 3rd, and 18th inter-base linkages are phosphodiester with remaining inter-base linkages being phosphorothioate, and all cytosine bases having a 5-methyl modification;
SEQ ID NO: 135 in a gapmer with a 12 base DNA central segment flanked by 2′-MOE RNA wings, in which 2d, 3rd, 4th, and 18th inter-base linkages are phosphodiester with remaining inter-base linkages being phosphorothioate, and all cytosine bases having a 5-methyl modification;
SEQ ID NO: 144 in a gapmer with a 12 base DNA central segment flanked by 2′-MOE RNA wings, in which 2d, 3rd, and 18th inter-base linkages are phosphodiester with remaining inter-base linkages being phosphorothioate, and all cytosine bases having a 5-methyl modification; and
SEQ ID NO: 144 in a gapmer with a 12 base DNA central segment flanked by 2′-MOE RNA wings, in which 2d, 3rd, 4th, and 18th inter-base linkages are phosphodiester with remaining inter-base linkages being phosphorothioate, and all cytosine bases having a 5-methyl modification.
24 . The composition of claim 23 , wherein the oligonucleotide knocks down expression of an SCN8A transcript to an amount about 50 to 70% when delivered to cells at about 1000 nM concentration compared to a control.
25 . A method comprising: administering to a subject with epilepsy a composition of claim 1 to thereby knockdown expression of a SCN8A gene.
26 . The method of claim 25 , wherein the epilepsy comprises Dravet syndrome, DEE13, or an epilepsy involving a pathogenic mechanism of excessive E/I balance.