Compositions and methods for delivery of RNA
Inventors
Khandhar, Amit • Reed, Steven • DUTHIE, Malcolm • Erasmus, Jesse • Carter, Darrick • BERUBE, Bryan J.
Assignees
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Publication Number
US-11679163-B2
Publication Date
2023-06-20
Expiration Date
Abstract
The disclosure provides nanoemulsion compositions and methods of making and using thereof to deliver a bioactive agent such as a nucleic acid to a subject. The nanoemulsion composition comprises a hydrophobic core based on inorganic nanoparticles in a lipid nanoparticle that allows imaging as well as delivering nucleic acids. Methods of using these particles for treatment and vaccination are also provided.
Core Innovation
The disclosed invention relates to a composition comprising RNA molecules and lipid nanoparticles formulated as a dry powder. The RNA molecules comprise a sequence encoding for an RNA polymerase complex region from an RNA virus and a protein antigen, and the RNA molecules are complexed to cationic lipids to form RNA-lipid nanoparticle complexes.
The lipid nanoparticles have a z-average diameter particle size measurement of about 20 nm to about 60 nm when measured using dynamic light scattering. The lipid nanoparticles comprise a surface comprising cationic lipids and a hydrophobic core comprising liquid oil, where lipids present in the hydrophobic core are in liquid phase at 25 degrees Celsius.
The disclosure further characterizes the hydrophobic core as being configured to include liquid oil and inorganic nanoparticles in some embodiments, and the inorganic core provides MRI-detectable reporter contrast for imaging and tracking. The described compositions also address RNA protection from RNases and support enhanced and long-lasting in vivo protein expression from RNA delivery.
In specific applications, the RNA payload includes replicon/self-replicating RNA and immune-stimulating RNAs, including mRNA and immunomodulating RNAs such as a RIG-I agonist PAMP and a TLR3 agonist Riboxxim. The document reports antigen activity, antibody induction, and immune activation following delivery in vivo, including results described for SARS-CoV-2 spike-encoding replicon RNA and vaccination/antibody responses in mice, rabbits, and nonhuman primates.
Claims Coverage
The partial content provides one independent claim (clm-00001). It defines a composition comprising RNA molecules encoding an RNA virus polymerase complex region and a protein antigen combined with lipid nanoparticles having a defined DLS-measured size range, a cationic lipid surface, and a liquid-oil hydrophobic core in liquid phase at 25 degrees Celsius, with RNA complexed to the cationic lipids, and the composition formulated as a dry powder. Inventive features are further refined in dependent claims by specifying particle size targets for RNA-lipid complexes, selection of specific surfactants, and inclusion of inorganic solid nanoparticles in the hydrophobic core.
RNA encoding an RNA polymerase complex region and a protein antigen
A composition comprising RNA molecules wherein the RNA molecules comprise a sequence encoding for an RNA polymerase complex region from an RNA virus and a protein antigen.
Lipid nanoparticles with a DLS-measured 20 nm to 60 nm z-average diameter
Lipid nanoparticles characterized as having a z-average diameter particle size measurement of about 20 nm to about 60 nm when measured using dynamic light scattering.
Cationic lipid surface and liquid-oil hydrophobic core in liquid phase at 25°C
The lipid nanoparticles comprise a surface comprising cationic lipids and a hydrophobic core comprising liquid oil, wherein lipids present in hydrophobic core are in liquid phase at 25 degrees Celsius.
RNA complexed to cationic lipids to form RNA-lipid nanoparticle complexes
The RNA molecules are complexed to the cationic lipids to form RNA-lipid nanoparticle complexes.
Dry powder formulation of the RNA-lipid nanoparticle complexes
The composition is formulated as a dry powder.
Optional refinement: RNA-lipid nanoparticle complexes sized at about 90 nm (DLS)
The composition is characterized by having RNA-lipid nanoparticle complexes having a z-average diameter of about 90 nm when measured using dynamic light scattering.
Optional refinement: hydrophobic surfactant selected from named sorbitan esters
The hydrophobic surfactant is selected from sorbitan monostearate, sorbitan monooleate, or sorbitan trioleate.
Optional refinement: inorganic solid nanoparticle in the hydrophobic core
The hydrophobic core includes a solid inorganic nanoparticle.
Across the provided claim set, coverage centers on an RNA-lipid nanoparticle composition formulated as a dry powder, where the RNA encodes an RNA virus RNA polymerase complex region and a protein antigen, and the lipid nanoparticles have a cationic lipid surface with a liquid-oil hydrophobic core in liquid phase at 25 degrees Celsius, complexing the RNA to form RNA-lipid nanoparticle complexes. Dependent refinements specify DLS-measured particle sizes for RNA-lipid complexes, selection of particular hydrophobic surfactants, and inclusion of solid inorganic nanoparticles in the hydrophobic core.
Stated Advantages
RNA protection from RNases.
Enhanced and long-lasting in vivo protein expression.
Retained antigen activity and robust antibody induction for a SARS-CoV-2 spike-encoding replicon RNA.
MRI reporter contrast for imaging and tracking, including enhanced T1/T2 relaxivity with iron-containing LIONs.
Immune activation from delivered immunomodulating RNAs, including induction of antibody responses described as Th1-biased neutralizing antibodies.
Documented Applications
Delivery of RNA (including replicon/self-replicating RNA and immune-stimulating RNAs) to induce protein expression in vivo.
Vaccination/antibody induction against SARS-CoV-2 spike using spike-encoding replicon RNA (repRNA-CoV2S), including antibody responses described in mice, rabbits, and nonhuman primates.
MRI imaging/tracking using inorganic core contrast from the described nanoemulsion-based LIONs (including iron-containing LIONs with enhanced relaxivity).
Immune activation using immunomodulating RNAs including a RIG-I agonist PAMP and a TLR3 agonist Riboxxim, with antibody expression described from antibody-encoding RNA.
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