Synergistic enhancement of the delivery of nucleic acids via blended formulations
Disclosed herein are pharmaceutical compositions that comprise “blends” of lipid nanoparticles and related methods of using such blended compositions to deliver polynucleotides to one or more target cells, tissues or organs. The blended compositions are generally characterized as being able to efficiently deliver polynucleotides to target cells and by their ability to enhance the expression of such polynucleotides and the production of functional proteins by target cells.
1. A method of delivering a messenger RNA (mRNA) to a subject, comprising administering to the subject a single pharmaceutical composition comprising a blend of at least a first and a second separately formed, non-identical lipid nanoparticles, each lipid nanoparticle comprising one or more PEG-modified lipids,
wherein the first lipid nanoparticle comprises a first mRNA and the second lipid nanoparticle comprises a second mRNA,
wherein the first lipid nanoparticle comprises a first cationic lipid and the second lipid nanoparticle comprises a second cationic lipid,
wherein the first cationic lipid and the second cationic lipid are non-identical, and
wherein an expression of a protein or a peptide encoded by the first mRNA or the second mRNA following the administration of the pharmaceutical composition to the subject exceeds the expression of the protein or the peptide encoded by the first mRNA or the second mRNA administered with the first lipid nanoparticle or the second lipid nanoparticle respectively, but without the other lipid nanoparticle by at least about two-fold.
2. The method of claim 1 , wherein the first cationic lipid is selected from the group consisting of C12-200, DOTAP (1,2-dioleyl-3-trimethylammonium propane), DODAP (1,2-dioleyl-3-dimethylammonium propane), DOTMA (1,2-di-O-octadecenyl-3-trimethylammonium propane), DLinDMA, DLin-KC2-DMA, HGT4003 and ICE.
3. The method of claim 1 , wherein the first lipid nanoparticle and the second lipid nanoparticle comprise one or more helper lipids.
4. The method of claim 3 , wherein the one or more helper lipids are selected from the group consisting of DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine), DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine), DOPE (1,2-dioleyl-sn-glycero-3-phosphoethanolamine), DPPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine), DMPE (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), DOPG (,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol)) and cholesterol.
5. The method of claim 1 , wherein the one or more PEG-modified lipids comprises a poly(ethylene)glycol chain of up to 5 kDa in length covalently attached to a lipid comprising one or more alkyl chains of C6-C20 in length.
6. The method of claim 1 , wherein the first lipid nanoparticle or the second lipid nanoparticle comprises one or more lipids selected from the group consisting of ICE, DSPC, CHOL, DODAP, DOTAP and C8-PEG-2000.
7. The method of claim 1 , wherein the first mRNA or the second mRNA encodes an enzyme.
8. The method of claim 1 , wherein the first mRNA or the second mRNA encodes a protein or a peptide, and the protein or the peptide encoded by the first mRNA or the second mRNA is secreted from one or more cells.
9. The method of claim 1 , wherein the first mRNA is selected from SEQ ID NO: 2 or SEQ ID NO: 3.
10. The method of claim 1 , wherein the first mRNA or the second mRNA comprises a chemical modification that renders the mRNA more stable.
11. The method of claim 1 , wherein the expression of the protein or the peptide encoded by the first mRNA following the administration of the pharmaceutical composition to the subject exceeds the expression of the protein or the peptide encoded by the first mRNA administered with the first lipid nanoparticle but without the second lipid nanoparticle by at least about five-fold, or by at least about ten-fold.
12. The method of claim 11 , wherein the first mRNA is the same as the second mRNA.
13. The method of claim 11 , wherein the first mRNA is different than the second mRNA.
14. The method of claim 1 , wherein the a ratio of the first lipid nanoparticle to the second lipid nanoparticle in the pharmaceutical composition is about 1:1 or about 2:1 or about 3:1 or about 4:1.