# US 10,946,107 B2 · Polynucleotide agents targeting hydroxyacid oxidase (glycolate oxidase, HAO1) and methods of use thereof

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- Number: US 10,946,107 B2
- Document ID: US10946107B2
- Status: Granted
- Kind: B2
- Application number: 15/841,486
- Filed: 2017-12-14
- Published: 2018-04-05
- Granted: 2021-03-16
- Earliest claimed: 2015-06-18
- Assignees: Alnylam Pharmaceuticals, Inc.
- Applicants: Alnylam Pharmaceuticals, Inc.
- Inventors: Gregory Hinkle
- Web page: https://patentgrep.com/patent/US10946107B2

## CPC

- A61K 48/005: {characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered}
- A61K 31/7115: Nucleic acids or oligonucleotides having modified bases, i.e. other than adenine, guanine, cytosine, uracil or thymine
- A61K 31/712: Nucleic acids or oligonucleotides having modified sugars, i.e. other than ribose or 2'-deoxyribose
- A61K 31/7125: Nucleic acids or oligonucleotides having modified internucleoside linkage, i.e. other than 3'-5' phosphodiesters
- A61P 13/02: of urine or of the urinary tract, e.g. urine acidifiers
- 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 15/1137: {against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138)}
- C12N 2310/11: Antisense
- 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 2310/3341: 5-Methylcytosine
- C12N 2310/341: Gapmers, i.e. of the type ===---===
- C12N 2310/345: having at least two different backbone modifications
- C12N 2310/346: having a combination of backbone and sugar modifications
- C12N 2310/3515: Lipophilic moiety, e.g. cholesterol
- C12Y 101/03015: (S)-2-Hydroxy-acid oxidase (1.1.3.15)
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12Y: ENZYMES

## Abstract

The invention relates to polynucleotide agents targeting an hydroxyacid oxidase (HAO1) gene, and methods of using such polynucleotide agents to inhibit expression of HAO1 and to treat subjects having an HAO1-associated disease, e.g., hyperoxaluria.

## Claims

### Claim 1

A single stranded antisense polynucleotide agent for inhibiting hydroxyacid oxidase (HAO1) expression, comprising at least 15 contiguous nucleotides of the nucleotide sequence 5′-GCACAGUGUCUCUUUGUCAA-3′ (SEQ ID NO:272), wherein the agent comprises
a gap segment consisting of linked deoxynucleotides;
a 5′-wing segment consisting of linked nucleotides;
a 3′-wing segment consisting of linked nucleotides;
wherein the gap segment is positioned between the 5′-wing segment and the 3′-wing segment and wherein each nucleotide of each wing segment comprises a modified sugar, and
wherein the agent is about 18 to about 24 nucleotides in length.

### Claim 2 (depends on 1)

The single stranded antisense polynucleotide agent of claim 1, wherein substantially all of the nucleotides are modified nucleotides.

### Claim 3 (depends on 1)

The single stranded antisense polynucleotide agent of claim 1, which is 20 nucleotides in length.

### Claim 4 (depends on 1)

The single stranded antisense polynucleotide agent of claim 1, wherein the gap segment is 5 to 14 2′-deoxynucleotides in length and each of the wing segments is 1 to 6 nucleotides in length.

### Claim 5 (depends on 1)

The single stranded antisense polynucleotide agent of claim 1, wherein the agent further comprises a ligand at the 3′-terminus.

### Claim 6 (depends on 5)

The single stranded antisense polynucleotide agent of claim 5, wherein the ligand is an N-acetylgalactosamine (GalNAc) derivative.

### Claim 7 (depends on 1)

A pharmaceutical composition for inhibiting expression of an hydroxyacid oxidase (HAO1) gene comprising the single stranded antisense polynucleotide agent of claim 1.

### Claim 8 (depends on 1)

A pharmaceutical composition comprising the single stranded antisense polynucleotide agent of claim 1, and a lipid formulation.

### Claim 9 (depends on 1, 7)

A method of inhibiting hydroxyacid oxidase (HAO1) expression in a cell, the method comprising:
(a) contacting the cell with the single stranded antisense polynucleotide agent of claim 1 or a pharmaceutical composition of claim 7; and
(b) maintaining the cell produced in step (a) for a time sufficient to obtain antisense inhibition of an HAO1 gene, thereby inhibiting expression of the HAO1 gene in the cell.

### Claim 10 (depends on 9)

The method of claim 9, wherein the cell is within a subject.

### Claim 11 (depends on 10)

The method of claim 10, wherein the subject is a human.

### Claim 12 (depends on 1, 7)

A method of treating a subject having a disease or disorder that would benefit from reduction in hydroxyacid oxidase (HAO1) expression, the method comprising administering to the subject a therapeutically effective amount of the single stranded antisense polynucleotide agent of claim 1 or a pharmaceutical composition of claim 7, thereby treating the subject.

### Claim 13 (depends on 1, 7)

A method of preventing at least one symptom in a subject having a disease or disorder that would benefit from reduction in hydroxyacid oxidase (HAO1) expression, the method comprising administering to the subject a prophylactically effective amount of the single stranded antisense polynucleotide agent of claim 1 or a pharmaceutical composition of claim 7, thereby preventing at least one symptom in the subject having a disorder that would benefit from reduction in HAO1 expression.

### Claim 14 (depends on 12, 13)

The method of claim 12 or 13, wherein the disorder is an hydroxyacid oxidase (HAO1)-associated disease.

### Claim 15 (depends on 14)

The method of claim 14, wherein the HAO1-associated disease is selected from the group consisting of primary hyperoxaluria and secondary hyperoxaluria.

### Claim 16 (depends on 1)

The single stranded antisense polynucleotide agent of claim 1, wherein the modified sugar moiety is selected from the group consisting of a 2′-O-methoxyethyl modified sugar moiety, a 2′-methoxy modified sugar moiety, a 2′-O-alkyl modified sugar moiety, and a bicyclic sugar moiety.

### Claim 17 (depends on 1)

The single stranded antisense polynucleotide agent of claim 1, further comprising a modified internucleoside linkage.

### Claim 18 (depends on 4)

The single stranded antisense polynucleotide agent of claim 4, wherein the 5′-wing segment is 4 to 6 nucleotides in length, the 3′-wing segment is 4 to 6 nucleotides in length, and the gap segment is 9 to 13 nucleotides in length.

### Claim 19 (depends on 7)

The pharmaceutical composition of claim 7, wherein the single stranded antisense polynucleotide agent is present in an unbuffered solution.

### Claim 20 (depends on 12, 13)

The method of claim 12 or 13, wherein the single stranded antisense polynucleotide agent is administered to the subject at a dose of about 0.01 mg/kg to about 10 mg/kg or about 0.5 mg/kg to about 50 mg/kg.

### Claim 21 (depends on 20)

The method of claim 20, wherein the single stranded antisense polynucleotide agent is administered to the subject once a week; twice a week; or twice a month.

### Claim 22 (depends on 12, 13)

The method of claim 12 or 13, wherein the single stranded antisense polynucleotide agent is administered to the subject subcutaneously.

### Claim 23 (depends on 1)

The single stranded antisense polynucleotide agent of claim 1, wherein all of the nucleotides are modified nucleotides.