# US 10,370,666 B2 · Ketohexokinase (KHK) iRNA compositions and methods of use thereof

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,370,666 B2
- Document ID: US10370666B2
- Status: Granted
- Kind: B2
- Application number: 15/224,805
- Filed: 2016-08-01
- Published: 2016-11-24
- Granted: 2019-08-06
- Earliest claimed: 2014-02-11
- Assignees: Alnylam Pharmaceuticals, Inc.
- Applicants: Alnylam Pharmaceuticals, Inc.
- Inventors: Kevin Fitzgerald; Brian Bettencourt; Gregory Hinkle; Jennifer Willoughby
- Web page: https://patentgrep.com/patent/US10370666B2

## CPC

- C12N 15/1137: {against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138)}
- A61K 31/713: Double-stranded nucleic acids or oligonucleotides
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 3/00: Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00)
- A61P 3/04: Anorexiants; Antiobesity agents
- A61P 3/06: Antihyperlipidemics
- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 9/12: Antihypertensives
- A61P 13/12: of the kidneys
- A61P 19/06: Antigout agents, e.g. antihyperuricemic or uricosuric agents
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 43/00: Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/315: Phosphorothioates
- C12N 2310/321: 2'-O-R Modification
- C12N 2310/322: 2'-R Modification
- C12N 2310/335: Modified T or U
- 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)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

The present invention relates to RNAi agents, e.g., double-stranded RNAi agents, targeting the ketohexokinase (KHK) gene, and methods of using such RNAi agents to inhibit expression of KHK and methods of treating subjects having a KHK-associated disorder, e.g., liver disease (e.g., fatty liver, steatohepatitis), dyslipidemia (e.g., hyperlipidemia, high LDL cholesterol, low HDL cholesterol, hypertriglyceridemia, postprandial hypertriglyceridemia), disorders of glycemic control (e.g., insulin resistance, diabetes), cardiovascular disease (e.g., hypertension, endothelial cell dysfunction), kidney disease (e.g., acute kidney disorder, tubular dysfunction, proinflammatory changes to the proximal tubules), metabolic syndrome, adipocyte dysfunction, visceral adipose deposition, obesity, hyperuricemia, gout, eating disorders, and excessive sugar craving.

## Claims

### Claim 1

A double stranded ribonucleic acid (dsRNA) agent for inhibiting expression of a ketohexokinase (KHK) gene,
wherein the dsRNA agent 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 3 nucleotides from the nucleotide sequence of nucleotides 798-833 of SEQ ID NO:5, and the antisense strand comprises at least 15 contiguous nucleotides of the corresponding nucleotide sequence of SEQ ID NO:6;
wherein substantially all of the nucleotides of the sense strand and substantially all of the nucleotides of the antisense strand are modified nucleotides, and
wherein the sense strand is conjugated to a ligand attached at the 3′-terminus.

### Claim 2 (depends on 1)

The dsRNA agent of claim 1, wherein the double stranded region is 17-25 nucleotide pairs in length.

### Claim 3 (depends on 2)

The dsRNA agent of claim 2, wherein the double stranded region is 19-23 nucleotide pairs in length.

### Claim 4 (depends on 1)

The dsRNA agent of claim 1, wherein each of the sense strand and the antisense strand independently is 21 to 23 nucleotides in length.

### Claim 5 (depends on 1)

The dsRNA agent of claim 1, wherein the sense strand is 21 nucleotides in length and the antisense strand is 23 nucleotides in length.

### Claim 6 (depends on 1)

The dsRNA agent of claim 1, wherein the modifications on the nucleotides are 2′-O-methyl or 2′-fluoro modifications.

### Claim 7 (depends on 1)

The dsRNA agent of claim 1, wherein the ligand is one or more GalNAc derivatives attached through a bivalent or trivalent branched linker.

### Claim 8 (depends on 1)

The dsRNA agent of claim 1, wherein the antisense strand comprises a nucleotide sequence selected from the group consisting of
(SEQ ID NO: 212)
5′-AUCUUUGCUGACAAACACCACGU-3′,
(SEQ ID NO: 211)
5′-AUUUGCUGACAAACACCACGUCU-3′,
(SEQ ID NO: 208)
5′-AAACACCACGUCUCCGUAGCCAA-3′,
(SEQ ID NO: 209)
5′-UAAACACCACGUCUCCGUAGCCA-3′,
(SEQ ID NO: 214)
5′-UACAUCUUUGCUGACAAACACCA-3′; and
(SEQ ID NO:213)
5′-ACAUCUUUGCUGACAAACACCAC-3′.

### Claim 9 (depends on 1)

The dsRNA agent of claim 1, wherein
(a) the sense strand comprises the nucleotide sequence
(SEQ ID NO: 162)
5′-GUGGUGUUUGUCAGCAAAGAU-3′
and the antisense strand comprises the nucleotide sequence
(SEQ ID NO: 212);
5′-AUCUUUGCUGACAAACACCACGU-3′
or
(b) the sense strand comprises the nucleotide sequence
(SEQ ID NO: 161)
5′-ACGUGGUGUUUGUCAGCAAAU-3′
and the antisense strand comprises the nucleotide sequence
(SEQ ID NO: 211);
5′-AUUUGCUGACAAACACCACGUCU-3′
or
(c) the sense strand comprises the nucleotide sequence
(SEQ ID NO: 158)
5′-GGCUACGGAGACGUGGUGUUU-3′
and the antisense strand comprises the nucleotide sequence
(SEQ ID NO: 208)
5′-AAACACCACGUCUCCGUAGCCAA-3′;
or
(d) the sense strand comprises the nucleotide sequence
(SEQ ID NO: 159)
5′-GCUACGGAGACGUGGUGUUUA-3′
and the antisense strand comprises the nucleotide sequence
(SEQ ID NO: 209)
5′-UAAACACCACGUCUCCGUAGCCA-3′;
or
(e) the sense strand comprises the nucleotide sequence
(SEQ ID NO: 164)
5′-GUGUUUGUCAGCAAAGAUGUA-3′
and the antisense strand comprises the nucleotide sequence
(SEQ ID NO: 214)
5′-UACAUCUUUGCUGACAAACACCA-3′;
or
(f) the sense strand comprises the nucleotide sequence
(SEQ ID NO: 163)
5′-GGUGUUUGUCAGCAAAGAUGU-3′
and the antisense strand comprises the nucleotide sequence
(SEQ ID NO: 213)
5′-ACAUCUUUGCUGACAAACACCAC-3′.

### Claim 10 (depends on 1)

The dsRNA agent of claim 1, wherein the dsRNA agent comprises:
(a) a sense strand consisting of the nucleotide sequence
(SEQ ID NO: 262)
5′-GfsusGfgUfgUfuUfGfUfcAfgCfaAfaGfaUfL96-3′
and an antisense strand consisting of the nucleotide sequence
(SEQ ID NO: 312)
5′-asUfscUfuUfgCfuGfacaAfaCfaCfcAfcsgsu-3′;
or
(b) a sense strand consisting of the nucleotide sequence
(SEQ ID NO: 261)
5′-AfscsGfuGfgUfgUfUfUfgUfcAfgCfaAfaUfL96-3′
and an antisense strand consisting of the nucleotide sequence
(SEQ ID NO: 311)
5′-asUfsuUfgCfuGfaCfaaaCfaCfcAfcGfuscsu-3′;
or
(c) a sense strand consisting of the nucleotide sequence
(SEQ ID NO: 258)
5′-GfsgsCfuAfcGfgAfGfAfcGfuGfgUfgUfuUfL96-3′
and an antisense strand consisting of the nucleotide sequence
(SEQ ID NO: 308)
5′-asAfsaCfaCfcAfcGfucuCfcGfuAfgCfcsasa-3′;
or
(d) a sense strand consisting of the nucleotide sequence
(SEQ ID NO: 259)
5′-GfscsUfaCfgGfaGfAfCfgUfgGfuGfuUfuAfL96-3′
and an antisense strand consisting of the nucleotide sequence
(SEQ ID NO: 309)
5′-usAfsaAfcAfcCfaCfgucUfcCfgUfaGfcscsa-3′;
or
(e) a sense strand consisting of the nucleotide sequence
(SEQ ID NO: 264)
5′-GfsusGfuUfuGfuCfAfGfcAfaAfgAfuGfuAfL96-3′
and an antisense strand consisting of the nucleotide sequence
(SEQ ID NO: 314)
5′- usAfscAfuCfuUfuGfcugAfcAfaAfcAfcscsa-3′;
or
(f) a sense strand consisting of the nucleotide sequence
(SEQ ID NO: 263)
5′-GfsgsUfgUfuUfgUfCfAfgCfaAfaGfaUfgUfL96-3′
and an antisense strand consisting of the nucleotide sequence
(SEQ ID NO: 313)
5′-asCfsaUfcUfuUfgCfugaCfaAfaCfaCfcsasc-3′,
wherein a, g, c, and u are 2′-O-methyl (2′-OMe) modified A, G, C and U nucleotides, respectively; Af, Gf, Cf and Uf are 2′ fluoro A, G, C and U modified nucleotides, respectively; s is a phosphorothioate linkage, and L96 is N-[tris(GalNAc-alkyl)-amidodecanoyl)]-4-hydroxyprolinol.

### Claim 11 (depends on 1)

The dsRNA agent of claim 1, wherein at least one strand comprises a 3′ overhang of at least 1 nucleotide; or a 3′ overhang of at least 2 nucleotides.

### Claim 12 (depends on 1)

The dsRNA agent of claim 1, wherein the double-stranded region is 15-30 nucleotide pairs in length.

### Claim 13 (depends on 1)

The dsRNA agent of claim 1, wherein each strand has 15-30 nucleotides.

### Claim 14

The dsRNA agent of 31, wherein the ligand is

### Claim 15 (depends on 14)

The dsRNA agent of claim 14, wherein the dsRNA agent is conjugated to the ligand as shown in the following schematic
wherein X is O or S.

### Claim 16 (depends on 1)

The dsRNA agent of claim 1, wherein said dsRNA agent further comprises at least one phosphorothioate or methylphosphonate internucleotide linkage.

### Claim 17 (depends on 1)

An isolated cell containing the dsRNA agent of claim 1.

### Claim 18 (depends on 1)

A pharmaceutical composition comprising the dsRNA agent of claim 1.