Oligonucleotide compounds comprising non-nucleotide overhangs
Inventors
Avkin-Nachum, Sharon • Feinstein, Elena
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-8859751-B2
Publication Date
2014-10-14
Expiration Date
Abstract
The invention relates to siRNA compounds comprising one non-nucleotide moiety covalently attached to at least one of the sense or antisense strands to down-regulate the expression of human genes. The invention also relates to pharmaceutical compositions comprising such compounds and a pharmaceutically acceptable carrier and to methods of treating and/or preventing the incidence or severity of various diseases or conditions associated with the target genes and/or symptoms associated with such diseases or conditions.
Core Innovation
The invention provides a double-stranded nucleic acid molecule comprising a sense strand and an antisense strand, in which the antisense strand comprises a non-nucleotide overhang covalently attached via a phosphodiester or phosphorothioate linkage to the 3′ or 2′ position of the sugar residue of the 3′ terminal nucleotide. The non-nucleotide overhang is selected from C3Pi, C3Ps, C3Pi-C3OH, C3Pi-C3Pi, C3Pi-C3Ps, C3Ps-C3OH, C3Ps-C3Pi, C3Ps-C3Ps, and related multi-linker and variant combinations, including abasic-related moieties.
The disclosed antisense overhangs include C3Pi-rAb, C3Pi-dAb, C3Ps-rAb, and C3Ps-dAb, with further listed variants such as rAbPi-C3OH, rAbPi-C3Pi, rAbPs-C3OH, rAbPs-C3Pi, dAbPi-C3OH, dAbPi-C3Pi, dAbPs-C3OH, and dAbPs-C3Pi. The disclosure further specifies that the sense strand may include a non-nucleotide overhang covalently attached to the 3′ terminal nucleotide, selected from propanol, a C3 alkyl moiety linked to a phosphodiester, or a C3 alkyl moiety linked to a phosphorothioate, or a pharmaceutically acceptable salt of such molecule.
The disclosed overhang-containing duplexes are Argonaute PAZ-recognized and are associated with improved stability/activity compared with blunt-ended and dTdT-ended controls. The disclosure further states reduced off-target effects and reduced immune response relative to blunt-ended and dTdT-ended controls, while supporting RNA interference through down-regulation of target gene expression.
Claims Coverage
The claim coverage centers on one independent claim and related dependent claims. The inventive features include an overhang-modified double-stranded nucleic acid, specific enumerated non-nucleotide overhang motifs, an optional sense-strand terminal overhang, and method language for down-regulating target gene expression.
Phosphodiester/phosphorothioate-linked antisense non-nucleotide overhang at 3′ or 2′ position
A double-stranded nucleic acid molecule comprising a sense strand and an antisense strand, wherein the antisense strand comprises a non-nucleotide overhang covalently attached via a phosphodiester or a phosphorothioate linkage to the 3′ or to the 2′ position of the sugar residue of the 3′ terminal nucleotide.
Enumerated C3Pi/C3Ps/C3OH and abasic overhang motifs
The non-nucleotide overhang is selected from C3Pi, C3Ps, C3Pi-C3OH, C3Pi-C3Pi, C3Pi-C3Ps, C3Ps-C3OH, C3Ps-C3Pi, C3Ps-C3Ps, C3Pi-C3Pi-C3OH, C3Ps-C3Ps-C3OH, C3Pi-C3Ps-C3OH, C3Ps-C3Pi-C3OH, C3Pi-C3Pi-C3Pi, C3Ps-C3Ps-C3Ps, C3Pi-C3Ps-C3Ps, C3Ps-C3Pi-C3Ps, C3Ps-C3Pi-C3Pi, C3Pi-C3Ps-C3Pi, C3Pi-rAb, C3Pi-dAb, C3Ps-rAb, C3Ps-dAb, rAbPi-C3OH, rAbPi-C3Pi, rAbPs-C3OH, rAbPs-C3Pi, dAbPi-C3OH, dAbPi-C3Pi, dAbPs-C3OH and dAbPs-C3Pi, wherein each Pi is a phosphodiester linkage and each Ps is a phosphorothioate linkage.
Optional sense-strand 3′ terminal overhang
A non-nucleotide overhang is covalently attached to the 3′ terminal nucleotide in the sense strand, where the overhang is selected from propanol, a C3 alkyl moiety linked to a phosphodiester, or a C3 alkyl moiety linked to a phosphorothioate.
Down-regulating target gene for disease treatment or prevention
A method of treating a subject with a disease or disorder linked to expression of a target gene by administering the molecule of claim 1, or a pharmaceutically acceptable salt, in an effective amount to down-regulate target gene expression.
The inventive concept is the antisense non-nucleotide overhang covalently attached at the sugar 3′ or 2′ position via phosphodiester/phosphorothioate linkages, with the overhang limited to enumerated C3Pi/C3Ps/C3OH and abasic-related variants. Dependent coverage further allows an additional sense-strand terminal overhang and includes method language for treating a subject by down-regulating target gene expression.
Stated Advantages
Improved stability/activity of the overhang-containing duplexes compared with blunt-ended and dTdT-ended controls.
Reduced off-target effects compared with blunt-ended and dTdT-ended controls.
Reduced immune response compared with blunt-ended and dTdT-ended controls.
Documented Applications
Therapeutic and/or prophylactic use for diseases or disorders linked to expression of a target gene, via down-regulation of target gene expression.
Hearing loss, glaucoma, ARDS, ischemia-reperfusion injury, ocular ischemic conditions, transplant-related injuries, AMD, and COPD.
Interested in licensing this patent?