Ice-based lipid nanoparticle formulation for delivery of mRNA
The present invention provides, among other things, compositions and methods of formulating nucleic acid-containing nanoparticles comprising no more than three distinct lipids components, one distinct lipid component being a sterol-based cationic lipid. In some embodiments, the present invention provides compositions and methods in which the lipid nanoparticles further comprise helper lipids and PEG-modified lipids. The resulting formulation comprises a high encapsulation percentage for nucleic acids.
1. A composition comprising:
nucleic acids; and
lipid nanoparticles encapsulating the nucleic acids, wherein each individual lipid nanoparticle comprises no more than three distinct lipid components, one distinct lipid component being a sterol-based cationic lipid comprising imidazole cholesterol ester (ICE),
wherein the sterol-based cationic lipid constitutes less than 70% molar ratio of the total lipids, and
further wherein the lipid nanoparticles have an encapsulation percentage for nucleic acids of at least 70%.
2. The composition of claim 1 , wherein the nucleic acids comprise mRNA encoding a Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein; and
lipid nanoparticles encapsulating the mRNA, wherein each individual lipid nanoparticle comprises no more than three distinct lipid components, one distinct lipid component being a sterol-based cationic lipid, and
wherein the lipid nanoparticles have an encapsulation percentage for mRNA of at least 80%.
3. The composition of claim 2 , wherein the mRNA encoding a CFTR protein comprises SEQ ID NO: 1, or a polynucleotide sequence at least 70%, 75%, 80%, 85%, 90%, or 95% identical to SEQ ID NO: 1.
4. A method of delivering mRNA in vivo comprising administering a composition of claim 2 to a subject, wherein the composition is administered intravenously, or by pulmonary delivery, wherein the pulmonary delivery comprises nebulization.
5. The method of claim 1 , wherein the nucleic acids are selected from DNA, siRNA, microRNA, and mRNA.
6. The composition of claim 1 , wherein the nucleic acids are mRNA encoding a protein or a peptide, wherein the mRNA is codon-optimized, and comprises one or more modified nucleotides.