Modulation of huntingtin expression
Inventors
Hung, Gene • Bennet, C. Frank • Freier, Susan M. • Kordasiewicz, Holly • Stanek, Lisa • Cleveland, Don W. • Cheng, Seng H. • Shihabuddin, Lamya
Assignees
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Publication Number
US-12297431-B2
Publication Date
2025-05-13
Expiration Date
Abstract
Provided herein are methods, compounds, and compositions for reducing expression of huntingtin mRNA and protein in an animal. Such methods, compounds, and compositions are useful to treat, prevent, delay, or ameliorate Huntington's disease, or a symptom thereof.
Core Innovation
The patent describes a compound comprising a single-stranded modified oligonucleotide designed as a gapmer. The gapmer has a defined architecture with a 5′ wing segment, a central gap segment, and a 3′ wing segment, with the wing segments formed by linked nucleosides and the central gap segment formed by linked deoxynucleosides. The nucleosides in each wing segment include a 2′-O-methoxyethyl sugar, and the oligonucleotide uses SEQ ID NO: 22 in which each cytosine is a 5-methylcytosine.
The compound further includes a defined internucleoside linkage pattern for the gapmer. Internucleoside linkages are either phosphorothioate internucleoside linkages or phosphodiester internucleoside linkages. Within the gap segment, as well as at the linkages connecting the central gap segment to the 5′ and 3′ wing segments, the internucleoside linkages are phosphorothioate, while the remaining internucleoside linkages are phosphodiester.
The documented biological content in the provided material connects this gapmer design to antisense oligonucleotide-mediated reduction of huntingtin mRNA. The summary content reports measurements of huntingtin mRNA reduction, including qRT-PCR and IC50/EC50-style readouts, and the experimental content spans in vitro and in vivo settings with human and animal huntingtin mRNA inhibition and downstream readouts including functional and toxicity markers.
The document also addresses in vivo delivery and duration-of-action through multiple administration contexts described in the provided material, including systemic tolerability after intraperitoneal dosing and CNS-focused dosing approaches such as intracerebroventricular delivery and intrathecal evaluation. Reported readouts include huntingtin mRNA inhibition and oligonucleotide concentrations in tissues, with associated tolerability measures including a microglial marker and liver-function transaminases and functional outcomes in animal models.
Claims Coverage
The partial content explicitly provides one independent claim directed to a gapmer oligonucleotide compound, with dependent claims reciting formulation components, salt forms, administration or therapeutic use to reduce huntingtin mRNA levels, and optional covalent linkage to a conjugate group. Inventive features focus on the gapmer architecture, the specified nucleobase sequence with 5-methylcytosine, the 2′-O-methoxyethyl sugars on the wing nucleosides, and a defined phosphorothioate/phosphodiester linkage pattern.
Gapmer architecture with defined 5′ wing, central gap, and 3′ wing segments
A single-stranded modified oligonucleotide that is a gapmer consisting of a 5′ wing segment, a central gap segment, and a 3′ wing segment, where the 5′ wing segment consists of five linked nucleosides, the central gap segment consists of ten linked deoxynucleosides, and the 3′ wing segment consists of five linked nucleosides.
Wing nucleosides with 2′-O-methoxyethyl sugar and SEQ ID NO: 22 base sequence with 5-methylcytosine
Each nucleoside of each wing segment comprises a 2′-O-methoxyethyl sugar, and the single-stranded modified oligonucleotide has the nucleobase sequence 5′-CTCAGTAACATTGACACCAC-3′ (SEQ ID NO: 22), wherein each cytosine is a 5-methylcytosine.
Defined phosphorothioate/phosphodiester linkage pattern for gap segment and termini
Each internucleoside linkage is either a phosphorothioate internucleoside linkage or a phosphodiester internucleoside linkage, where each internucleoside linkage within the gap segment is phosphorothioate, the internucleoside linkage connecting the central gap segment to the 5′ wing segment is phosphorothioate, the internucleoside linkage connecting the central gap segment to the 3′ wing segment is phosphorothioate, the 5′-most internucleoside linkage of the 5′ wing segment is phosphorothioate, the 3′-most internucleoside linkage of the 3′ wing segment is phosphorothioate, and the remaining internucleoside linkages are phosphodiester.
Pharmaceutical composition with specified formulation components and optional salt forms
Dependent coverage recites a pharmaceutical composition that comprises the compound with pharmaceutically acceptable carrier or diluent, or that consists essentially of the compound and PBS, and further specifies that the pharmaceutical composition can use a pharmaceutically acceptable salt that is either a sodium salt or a potassium salt.
Reducing huntingtin mRNA levels in an animal by administering a therapeutically effective amount
Dependent coverage recites a method reducing huntingtin mRNA levels in an animal by administering a therapeutically effective amount of the pharmaceutical composition.
Covalent linkage of the single-stranded modified oligonucleotide to a conjugate group
Dependent coverage further recites that the single-stranded modified oligonucleotide is covalently linked to a conjugate group.
Overall, the claim set presented focuses on a specific gapmer antisense oligonucleotide compound defined by the 5′ wing/central gap/3′ wing architecture, 2′-O-methoxyethyl sugars on wing nucleosides with a defined SEQ ID NO: 22 sequence containing 5-methylcytosine, and a specific phosphorothioate/phosphodiester internucleoside linkage pattern emphasizing phosphorothioate linkages in and around the central gap and at the termini. Dependent claims add pharmaceutical composition formulation elements, optional salt specification, methods of reducing huntingtin mRNA levels in an animal, and an optional covalent linkage to a conjugate group.
Stated Advantages
Reduces huntingtin mRNA levels in an animal.
Supports antisense gapmer targeting of huntingtin expression for Huntington’s disease contexts.
Documented Applications
Reducing huntingtin mRNA levels in an animal, including Huntington’s disease.
Evaluation involving in vivo and in vitro assessment of huntingtin mRNA knockdown and related huntingtin protein/phenotypes.
Use of intracerebroventricular infusion/bolus and systemic administration routes in described evaluation contexts.
Biological readouts using motor and behavioral assays and cellular readouts including transaminases (ALT, AST) in disclosed contexts.
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