Compositions and methods for delivery of RNA

Inventors

Khandhar, Amit • Reed, Steven • DUTHIE, Malcolm • Erasmus, Jesse • Carter, Darrick • BERUBE, Bryan J.

Assignees

HDT Bio Corp

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 invention relates to lipid nanoparticles forming nucleic acid-lipid nanoparticle complexes, where nucleic acids include sequences encoding for a protein region and are complexed to cationic lipids. The lipid nanoparticles are characterized by a z-average diameter particle size measurement in a range of 20 nm to about 60 nm when measured using dynamic light scattering. A surface comprising cationic lipids and a hydrophobic core comprising liquid oil are used such that lipids present in the hydrophobic core are in liquid phase at 25 degrees Celsius.

The compositions further include an adjuvant for innate immune stimulation. The nucleic acids are protected in the nanoparticle complexes such that the RNA is protected from RNases and enables intracellular protein expression. The disclosed immune-stimulation concepts include immune responses such as vaccination and antibody induction, including Th1 bias and T-cell responses.

In the disclosed compositions, lipid-inorganic nanoparticle (LION) nanoemulsion concepts are described for RNA delivery, combining a hydrophobic oil phase containing squalene and terpenoids with an inorganic MRI-detectable core such as iron oxide SPIO or aluminum hydroxide/oxyhydroxide nanoparticles. The RNA payload can include mRNA and self-replicating RNA/replicons, including immune-stimulatory RNAs such as RIG-I agonists or TLR3 agonists. MRI/relaxivity-based imaging and tracking concepts are described alongside nanoparticle stability, reporter assays for innate immune activation, and in vivo immunogenicity across multiple mammalian models.

Claims Coverage

The independent claim coverage centers on a composition that combines nucleic acids encoding a protein region with lipid nanoparticles having defined cationic-lipid surface and liquid-phase hydrophobic oil core properties, specifies dynamic light scattering z-average particle size constraints, requires nucleic acids complexed to the cationic lipids, and includes an adjuvant for innate immune stimulation. Main inventive features are further refined in dependent claims by specifying RNA type and particular adjuvant and lipid identities, along with additional formulation constituents and more specific particle-size constraints.

Overall, the claim coverage ties together nucleic acids encoding a protein region complexed to cationic lipids in lipid nanoparticles with defined dynamic light scattering z-average size and a liquid-phase hydrophobic oil core at 25 degrees Celsius, and requires an innate immune stimulation adjuvant, with dependent claim refinements specifying TLR or RIG-I agonists, an mRNA payload, and particular cationic lipids and additional constituents such as squalene, sorbitan monostearate, and polysorbate 80.

Stated Advantages

Documented Applications