Reduced size self-delivering RNAI compounds
Inventors
Khvorova, Anastasia • Salomon, William • Kamens, Joanne • Samarsky, Dmitry • Woolf, Tod M. • Cardia, James
Assignees
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Publication Number
US-11118178-B2
Publication Date
2021-09-14
Expiration Date
Abstract
The present invention relates to methods for in vivo administration of sd-rxRNA molecules.
Core Innovation
The invention provides reduced size self-delivering RNAi (sd-rxRNA®) molecules as isolated asymmetric minimally sized RNAi triggers for inhibiting expression of a target gene in a mammal. The sd-rxRNA® comprises a guide strand and a passenger strand, with a double stranded region of 8–15 nucleotides and a single stranded region at the 3′ end of the guide strand of 4–12 nucleotides.
The single stranded region contains phosphorothioate modifications, with at least 40% of the nucleotides modified and, in some content, at least 60% of the nucleotides modified. At least two Us and/or Cs include a hydrophobic modification comprising an octyl or a pyridyl amide modification.
The disclosure further describes hydrophobic substituents attached to an oligonucleotide or nucleic acid, including cholesterol, lipophilic vitamins, vitamin A, vitamin E, steroid-type hydrophobic groups, sterol conjugates, cholesterol conjugates, 2′-modifications, chimeric DNA/RNA constructs, particles, and delivery vehicles.
Claims Coverage
The consolidated claim coverage centers on one independent method claim for inhibiting expression of a target gene in a mammal using sd-rxRNA® with a defined reduced-size guide/passenger strand architecture and specific nucleic-acid modification patterns. The inventive features include the strand architecture, phosphorothioate-modified single stranded region, an overall modified-nucleotide threshold, and hydrophobic U/C base modifications; dependent refinements add higher modification fractions, additional conjugation and delivery context limitations.
Administering sd-rxRNA® to inhibit target gene expression in a mammal
A method for inhibiting expression of a target gene of a mammal by administering an sd-rxRNA® in an effective amount.
Reduced-size double stranded and single stranded regions with defined lengths
The sd-rxRNA® includes a guide strand and a passenger strand, with a double stranded region from 8–15 nucleotides long and a single stranded region at the 3′ end of the guide strand that is 4–12 nucleotides long.
Single stranded region phosphorothioate modification pattern with at least 40% modified nucleotides
The single stranded region contains 3 to 12 phosphorothioate modifications, and at least 40% of the nucleotides are modified.
Hydrophobic U/C modifications using octyl or pyridyl amide
At least two Us and/or Cs include a hydrophobic modification comprising an octyl or a pyridyl amide modification.
Higher nucleic-acid modification fraction
The method is performed such that at least 60% of the nucleotides are modified.
Specific administration routes
The sd-rxRNA is administered subcutaneously, intravenously, or via intrathecal delivery.
3′-linked cholesterol and/or increased phosphorothioate count
The sd-rxRNA is 3′-linked to cholesterol and/or the guide strand includes at least 5 phosphorothioate modifications.
Tumor delivery
Delivering sd-rxRNA to a tumor, optionally via intravenous administration or via direct injection into the tumor.
Overall, the claims focus on administering sd-rxRNA® molecules having a specific guide/passenger strand architecture together with phosphorothioate modification, a defined modified-nucleotide fraction, and hydrophobic octyl or pyridyl amide modifications. Dependent claims further specify higher modification fractions, optional cholesterol conjugation, particular delivery routes, and tumor delivery context.
Stated Advantages
Increased serum stability.
Systemic tissue distribution without a delivery vehicle.
Effective delivery to liver, heart, lung/alveolar macrophages, and brain/spinal cord.
Increased serum half-life greater than 12 h.
Compatibility with repetitive dosing and continuous infusion.
Improved liver uptake and efficacy for repetitive dosing.
Route-dependent clearance/retention.
Documented Applications
Treating diseases by targeting genes including VEGF/VEGFR, HER2, PCSK9, ApoB, MAP4K4, and PPIB.
Delivering sd-rxRNA to a tumor, including optional intravenous administration or direct injection into the tumor.
Lung targeting via insufflation or intratracheal spray delivery context.
In vivo delivery and tissue distribution using systemic administration via IV, subcutaneous administration, and insufflation, including targeting to lungs preferentially to alveolar macrophages.
Targeting and delivery contexts including liver, spleen, heart, skin, lungs/alveolar macrophages, and tumor delivery.
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