# US 11,993,774 B2 · Huntingtin (HTT) iRNA agent compositions and methods of use thereof

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- Number: US 11,993,774 B2
- Document ID: US11993774B2
- Status: Granted
- Kind: B2
- Application number: 18/327,139
- Filed: 2023-06-01
- Published: 2023-12-07
- Granted: 2024-05-28
- Earliest claimed: 2021-03-29
- Assignees: Alnylam Pharmaceuticals, Inc.
- Applicants: Alnylam Pharmaceuticals, Inc.
- Inventors: Mangala Meenakshi Soundarapandian; James D. McIninch; Mark K. Schlegel; Adam Castoreno
- Web page: https://patentgrep.com/patent/US11993774B2

## CPC

- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/312: Phosphonates
- C12N 2310/315: Phosphorothioates
- C12N 2310/321: 2'-O-R Modification
- C12N 2310/322: 2'-R Modification
- C12N 2310/323: modified ring structure
- C12N 2310/346: having a combination of backbone and sugar modifications
- C12N 2310/3515: Lipophilic moiety, e.g. cholesterol
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

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.

## Claims

### Claim 1

A double stranded ribonucleic acid (dsRNA) agent, or a pharmaceutically acceptable salt thereof, for inhibiting expression of Huntingtin (HTT) in a cell, comprising 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 (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.

### Claim 2 (depends on 1)

The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 3 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 3 nucleotides from the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79.

### Claim 3 (depends on 1)

The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 2 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 2 nucleotides from the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79.

### Claim 4 (depends on 1)

The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, wherein the sense strand comprises at least 15 contiguous nucleotides differing by no more than 1 nucleotide 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 1 nucleotide from the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79.

### Claim 5 (depends on 1)

The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, 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.

### Claim 6 (depends on 1)

The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, 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.

### Claim 7 (depends on 1)

The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, wherein the sense strand comprises at least 17 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 17 contiguous nucleotides differing by no more than 4 nucleotides from the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79.

### Claim 8 (depends on 1)

The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, wherein the sense strand comprises at least 18 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 18 contiguous nucleotides differing by no more than 4 nucleotides from the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79.

### Claim 9 (depends on 1)

The dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 1, wherein the sense strand comprises at least 19 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 19 contiguous nucleotides differing by no more than 4 nucleotides from the nucleotide sequence 5′-VPusAfsucdAg(C2p)uuuudCcAfgdGgucgcscsg-3′ of SEQ ID NO:79.

### Claim 10 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9)

An isolated cell containing the dsRNA agent, or a pharmaceutically acceptable salt thereof, of any one of claims 1-9.

### Claim 11 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9)

A pharmaceutical composition for inhibiting expression of a gene encoding HTT, comprising the dsRNA agent, or a pharmaceutically acceptable salt thereof, of any one of claims 1-9.

### Claim 12 (depends on 11)

The pharmaceutical composition of claim 11, wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in an unbuffered solution.

### Claim 13 (depends on 11)

The pharmaceutical composition of claim 11, wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a buffer solution.

### Claim 14

A double stranded ribonucleic acid (dsRNA) agent, or a pharmaceutically acceptable salt thereof, for inhibiting expression of Huntingtin (HTT) in a cell, comprising 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.

### Claim 15 (depends on 14)

An isolated cell containing the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 14.

### Claim 16 (depends on 15)

A pharmaceutical composition for inhibiting expression of a gene encoding HTT, comprising the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 15.

### Claim 17 (depends on 16)

The pharmaceutical composition of claim 16, wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in an unbuffered solution.

### Claim 18 (depends on 17)

The pharmaceutical composition of claim 17, wherein the unbuffered solution is saline or water.

### Claim 19 (depends on 16)

The pharmaceutical composition of claim 16, wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a buffer solution.

### Claim 20 (depends on 19)

The pharmaceutical composition of claim 19, wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof.

### Claim 21 (depends on 19)

The pharmaceutical composition of claim 19, wherein the buffer solution is phosphate buffered saline (PBS).

### Claim 22

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.

### Claim 23 (depends on 22)

An isolated cell containing the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 22.

### Claim 24 (depends on 22)

A pharmaceutical composition for inhibiting expression of a gene encoding HTT, comprising the dsRNA agent, or a pharmaceutically acceptable salt thereof, of claim 22.

### Claim 25 (depends on 24)

The pharmaceutical composition of claim 24, wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in an unbuffered solution.

### Claim 26 (depends on 25)

The pharmaceutical composition of claim 25, wherein the unbuffered solution is saline or water.

### Claim 27 (depends on 24)

The pharmaceutical composition of claim 24, wherein the dsRNA agent, or a pharmaceutically acceptable salt thereof, is present in a buffer solution.

### Claim 28 (depends on 27)

The pharmaceutical composition of claim 27, wherein the buffer solution comprises acetate, citrate, prolamine, carbonate, or phosphate or any combination thereof.

### Claim 29 (depends on 27)

The pharmaceutical composition of claim 27, wherein the buffer solution is phosphate buffered saline (PBS).

### Claim 30

A sodium salt of a double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of Huntingtin (HTT) in a cell,
wherein the dsRNA agent 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.