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 pharmaceutical composition for delivery of messenger RNA (mRNA) to a cell, wherein the composition is a single pharmaceutical composition comprising a blend of at least a first lipid nanoparticle and a second separately formed, non-identical lipid nanoparticle,
wherein each lipid nanoparticle comprises one or more PEG-modified lipids,
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;
wherein the first lipid nanoparticle encapsulates a first mRNA and the second lipid nanoparticle encapsulates a second mRNA;
wherein the first lipid nanoparticle and the second lipid nanoparticle in the pharmaceutical composition are in a ratio of at least about 2:1 based on the mass of the encapsulated mRNAs; and
wherein expression of a protein or a peptide encoded by the first mRNA or the second mRNA following the delivery of the pharmaceutical composition to the cell 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, and
wherein the amount of the mRNA required to effectuate a therapeutic effect following the delivery of the pharmaceutical composition to the cell is reduced by at least about two-fold relative to the amount of mRNA required to effectuate a therapeutic effect following the delivery of 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.
2. The pharmaceutical composition of claim 1 , wherein the first lipid nanoparticle and the second lipid nanoparticle comprise one or more helper lipids.
3. The pharmaceutical composition 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 C 6 -C 20 in length.
4. The pharmaceutical composition of claim 1 , wherein the first lipid nanoparticle comprises
one or more PEG-modified lipids that 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 C 6 -C 20 in length; and
a helper lipid 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; and
the second lipid nanoparticle comprises
one or more PEG-modified lipids that 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 C 6 -C 20 in length; and
a helper lipid 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 pharmaceutical composition of claim 1 , wherein the mRNA comprises a chemical modification.
6. The pharmaceutical composition of claim 1 , wherein the expression of the mRNA following the delivery of the pharmaceutical composition to the cell exceeds the expression of the mRNA administered with the first lipid nanoparticle but without the second lipid nanoparticle by at least about five-fold.
7. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition is delivered to the cell in vivo.
8. The pharmaceutical composition of claim 1 , wherein the ratio of the first lipid nanoparticle to the second lipid nanoparticle in the pharmaceutical composition is about 2:1.
9. The pharmaceutical composition of claim 1 , wherein the ratio of the first lipid nanoparticle to the second lipid nanoparticle in the pharmaceutical composition is about 3:1.
10. The pharmaceutical composition of claim 1 , wherein the ratio of the first lipid nanoparticle to the second lipid nanoparticle in the pharmaceutical composition is about 4:1.
11. A pharmaceutical composition for delivery of a messenger RNA (mRNA) to one or more target cells, said composition comprising a blend of at least a first lipid nanoparticle and a second separately formed, non-identical lipid nanoparticle,
wherein each lipid nanoparticle comprises one or more PEG-modified lipids,
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 the first lipid nanoparticle encapsulates an mRNA and the second lipid nanoparticle does not encapsulate any polynucleotides; and
wherein the first lipid nanoparticle and the second lipid nanoparticle in the pharmaceutical composition are in a ratio of at least about 2:1 based on the mass of the encapsulated mRNA; and
wherein expression of a protein or a peptide encoded by the mRNA following the delivery of the pharmaceutical composition to the cell exceeds expression of the protein or peptide encoded by the otherwise identical amount of mRNA administered with the first lipid nanoparticle but without the second lipid nanoparticle.
12. The pharmaceutical composition of claim 11 , wherein the first lipid nanoparticle and the second lipid nanoparticle comprise one or more helper lipids.
13. The pharmaceutical composition of claim 11 , 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 C 6 -C 20 in length.
14. The pharmaceutical composition of claim 11 , wherein the first lipid nanoparticle comprises
one or more PEG-modified lipids that 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 C 6 -C 20 in length; and
a helper lipid 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; and
the second lipid nanoparticle comprises
one or more PEG-modified lipids that 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 C 6 -C 20 in length; and
a helper lipid 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.
15. The pharmaceutical composition of claim 11 , wherein the lipid nanoparticle that does not encapsulate any polynucleotides is a C12-200-based cationic lipid nanoparticle.
16. The pharmaceutical composition of claim 11 , wherein the ratio of the first lipid nanoparticle to the second lipid nanoparticle in the pharmaceutical composition is about 2:1.
17. The pharmaceutical composition of claim 11 , wherein the ratio of the first lipid nanoparticle to the second lipid nanoparticle in the pharmaceutical composition is about 3:1.
18. The pharmaceutical composition of claim 11 , wherein the ratio of the first lipid nanoparticle to the second lipid nanoparticle in the pharmaceutical composition is about 4:1.
19. The pharmaceutical composition of claim 11 , wherein the ratio of the first lipid nanoparticle to the second lipid nanoparticle in the pharmaceutical composition is about 5:1.
20. The pharmaceutical composition of claim 1 , wherein the ratio of the first lipid nanoparticle to the second lipid nanoparticle in the pharmaceutical composition is about 5:1.