HUNTINGTIN (HTT) IRNA AGENT COMPOSITIONS AND METHODS OF USE THEREOF
The disclosure relates to double stranded ribonucleic acid (dsRNAi) agents and compositions targeting a Huntingtin (HTT) gene, as well as methods of inhibiting expression of an HTT gene and methods of treating subjects having an HTT-associated disease or disorder, e.g., Huntington's disease, using such dsRNAi agents and compositions.
1 . A method of inhibiting expression of a huntingtin (HTT) gene in a cell, the method comprising:
(a) contacting the cell with a double stranded ribonucleic acid (dsRNA) agent, or a pharmaceutically acceptable salt thereof, for inhibiting expression of Huntingtin (HTT) in a cell,
wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 4 nucleotides from the nucleotide sequence 5′-gscsgac(Chd)CfuGfGfAfaaagcugasusa-3′ of SEQ ID NO:65 and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 4 nucleotides from the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; s is a phosphorothioate linkage; Af, Cf, and Gf are 2′-fluoro A, C, and G, respectively; (Chd) is 2′-O-hexadecyl-cytosine-3′-phosphate; C2p is cytidine-2′-phosphate; dA, dG, and dC are 2′-deoxy A, G, and C, respectively; and VP is 5′-vinyl phosphonate; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain degradation of the mRNA transcript of the HTT gene, thereby inhibiting expression of the HTT gene in the cell.
2 . The method of claim 1 , wherein the cell is within a subject.
3 . The method of claim 2 , wherein the subject is a human.
4 . The method of claim 2 , wherein the subject has been diagnosed with an HTT-associated disease.
5 . The method of claim 4 , wherein the HTT-associated disease is Huntington's disease.
6 . A method of treating a subject diagnosed with an HTT-associated disease, the method comprising administering to the subject a therapeutically effective amount of a double stranded ribonucleic acid (dsRNA) agent, or a pharmaceutically acceptable salt thereof, for inhibiting expression of Huntingtin (HTT) in a cell,
wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 4 nucleotides from the nucleotide sequence 5′-gscsgac(Chd)CfuGfGfAfaaagcugasusa-3′ of SEQ ID NO:65 and the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 4 nucleotides from the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; s is a phosphorothioate linkage; Af, Cf, and Gf are 2′-fluoro A, C, and G, respectively; (Chd) is 2′-O-hexadecyl-cytosine-3′-phosphate; C2p is cytidine-2′-phosphate; dA, dG, and dC are 2′-deoxy A, G, and C, respectively; and VP is 5′-vinyl phosphonate, thereby treating the subject.
7 . The method of claim 6 , wherein the HTT-associated disease is Huntington's disease.
8 . The method of claim 6 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is administered to the subject intrathecally.
9 . The method of claim 6 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is administered to the subject intravenously.
10 . The method of claim 6 , further comprising administering to the subject an additional agent suitable for treatment or prevention of an HTT-associated disorder.
11 . The method of claim 6 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a pharmaceutical composition.
12 . The method of claim 11 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in an unbuffered solution.
13 . The method of claim 11 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a buffer solution.
14 . A method of treating a subject diagnosed with an HTT-associated disease, the method comprising administering to the subject a therapeutically effective amount of a double stranded ribonucleic acid (dsRNA) agent, or a pharmaceutically acceptable salt thereof, for inhibiting expression of Huntingtin (HTT) in a cell,
wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand comprises the nucleotide sequence 5′-gscsgac(Chd)CfuGfGfAfaaagcugasusa-3′ of SEQ ID NO:65 and the antisense strand comprises the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; s is a phosphorothioate linkage; Af, Cf, and Gf are 2′-fluoro A, C, and G, respectively; (Chd) is 2′-O-hexadecyl-cytosine-3′-phosphate; C2p is cytidine-2′-phosphate; dA, dG and dC are 2′-deoxy A, G, and C, respectively; and VP is 5′-vinyl phosphonate, thereby treating the subject.
15 . The method of claim 14 , wherein the HTT-associated disease is Huntington's disease.
16 . The method of claim 14 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is administered to the subject intrathecally.
17 . The method of claim 14 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is administered to the subject intravenously.
18 . The method of claim 14 , further comprising administering to the subject an additional agent suitable for treatment or prevention of an HTT-associated disorder.
19 . The method of claim 14 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a pharmaceutical composition.
20 . The method of claim 19 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in an unbuffered solution.
21 . The method of claim 19 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a buffer solution.
22 . The method of claim 19 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a sodium salt form.
23 . A method of treating a subject diagnosed with an HTT-associated disease, the method comprising administering to the subject a therapeutically effective amount of a double stranded ribonucleic acid (dsRNA) agent, or a pharmaceutically acceptable salt thereof, for inhibiting expression of Huntingtin (HTT) in a cell,
wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, comprises a sense strand and an antisense strand forming a double stranded region,
wherein the sense strand consists of the nucleotide sequence 5′-gscsgac(Chd)CfuGfGfAfaaagcugasusa-3′ of SEQ ID NO:65 and the antisense strand consists of the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; s is a phosphorothioate linkage; Af, Cf, and Gf are 2′-fluoro A, C, and G, respectively; (Chd) is 2′-O-hexadecyl-cytosine-3′-phosphate; C2p is cytidine-2′-phosphate; dA, dG and dC are 2′-deoxy A, G, and C, respectively; and VP is 5′-vinyl phosphonate, thereby treating the subject.
24 . The method of claim 23 , wherein the HTT-associated disease is Huntington's disease.
25 . The method of claim 23 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is administered to the subject intrathecally.
26 . The method of claim 23 , wherein the dsRNA agent is administered to the subject intravenously.
27 . The method of claim 23 , further comprising administering to the subject an additional agent suitable for treatment or prevention of an HTT-associated disorder.
28 . The method of claim 23 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a pharmaceutical composition.
29 . The method of claim 28 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in an unbuffered solution.
30 . The method of claim 29 , wherein the unbuffered solution is saline or water.
31 . The method of claim 28 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a buffer solution.
32 . The method of claim 31 , wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof.
33 . The method of claim 31 , wherein the buffer solution is phosphate buffered saline (PBS).
34 . The method of claim 28 , wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a sodium salt form.
35 . A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of Huntingtin (HTT) in a cell, or a pharmaceutically acceptable salt thereof, wherein the dsRNA comprises a sense strand and an antisense strand forming a double stranded region, wherein:
(a) the antisense strand comprises at least 15 contiguous nucleotides differing by no more than 3 nucleotides from any one of the antisense nucleotide sequences in any one of Tables 2-5; or
(b) wherein the sense strand comprises the nucleotide sequence 5′-gscsgac(Chd)CfuGfGfAfaaagcugasusa-3′ of SEQ ID NO:65 and the antisense strand comprises the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79,
wherein a, g, c and u are 2′-O-methyl (2′-OMe) A, G, C, and U, respectively; s is a phosphorothioate linkage; Af, Cf, and Gf are 2′-fluoro A, C, and G, respectively; (Chd) is 2′-O-hexadecyl-cytosine-3′-phosphate; C2p is cytidine-2′-phosphate; dA, dG and dC are 2′-deoxy A, G, and C, respectively; and VP is 5′-vinyl phosphonate.