RNA formulations
This disclosure provides improved lipid-based compositions, including lipid nanoparticle compositions, and methods of use thereof for delivering agents in vivo including nucleic acids and proteins.
1. A composition comprising an enriched population of lipid nanoparticles (LNPs), wherein the LNPs have an outer shell and an inner core and comprise an ionizable lipid, a phospholipid, a poly(ethylene glycol) (PEG) lipid, wherein at least 50% of the LNPs comprise mRNA encapsulated within the inner core and wherein the outer shell comprises at least 95% of the total PEG lipid in the LNP, and wherein the LNP's are formed by a nanoprecipitation process including a step in which additional PEG lipid was added to the LNP's after an initial nanoprecipitation reaction.
2. The composition of claim 1 , further comprising a structural lipid.
3. The composition of claim 2 , wherein at least 50% of the LNPs have a ratio of ionizable lipid:phospholipid:structural lipid of 35-50:30-40: 20-30.
4. The composition of claim 2 , wherein:
(a) the LNP is insensitive to accelerated blood clearance upon repeated administration in vivo within 10 days;
(b) at least 50% of the LNPs have 1-5 inner shells;
(c) the structural lipid is cholesterol;
(d) 10-30% of the LNP, exclusive of mRNA, is the structural lipid; and/or
(e) the outer shell is comprised of 10-30% of the structural lipid.
5. The composition of claim 1 , wherein at least 50% of the LNPs have an outer shell fluidity value of greater than a threshold polarization level.
6. The composition of claim 1 , wherein a quantitative value of the amount of mRNA encapsulated in the LNP is generated using an ion-exchange (IEX) chromatography assay.
7. The composition of claim 1 , wherein:
(a) at least about 50% of the LNPs in the population have mRNA encapsulated therein, as determined by ion-exchange chromatography (IEX); and/or
(b) the LNPs have an encapsulation efficiency of at least about 50% as determined by ion-exchange chromatography (IEX).
8. The composition of claim 1 , wherein the phospholipid is DSPC.
9. A composition comprising a population of lipid nanoparticles (LNPs), the LNPs comprise an ionizable amino lipid, a poly(ethylene glycol) (PEG) lipid, a phospholipid, wherein:
(a) at least about 50% of the LNPs in the population have mRNA encapsulated therein, as determined by ion-exchange chromatography (IEX), and wherein the LNP's are formed by a nanoprecipitation process including a step in which additional PEG lipid was added to the LNP's after an initial nanoprecipitation reaction; and/or
(b) the LNPs have an encapsulation efficiency of at least about 50% as determined by ion-exchange chromatography (IEX).
10. The composition of claim 9 , further comprising a structural lipid.
11. The composition of claim 10 , wherein the structural lipid is cholesterol or a cholesterol derivative.
12. The composition of claim 9 , wherein the composition is enriched for LNPs:
(a) lacking B1a cell-stimulating phospholipid epitopes, and/or
(b) lacking scavenger receptor ligands.
13. The composition of claim 12 , wherein at least about 50% of the LNPs:
(a) lack B1a cell-stimulating phospholipid epitopes, and/or
(b) lack scavenger receptor ligands.
14. The composition of claim 9 , wherein the composition is enriched for LNPs having a majority of the total phospholipid present in the outer LNP shell.
15. The composition of claim 14 , wherein at least about 50% of the LNPs have the majority of the total phospholipid present in the outer LNP shell.
16. The composition of claim 9 , wherein the composition is enriched for LNPs having more than 50% of the total phospholipid present on the surface.
17. The composition of claim 16 , wherein at least about 50% of the LNPs have more than 50% of the total phospholipid present on the surface.
18. The composition of claim 9 , wherein the composition is enriched for LNPs having a majority of the total PEG lipid present in the outer LNP shell.
19. The composition of claim 18 , wherein at least about 50% of the LNPs have the majority of the total PEG lipid present in the outer LNP shell.
20. The composition of claim 9 , wherein the composition is enriched for LNPs having more than 50% of the total PEG lipid present on the surface.
21. The composition of claim 20 , wherein at least about 50% of the LNPs have more than 50% of the total PEG lipid present on the surface.
22. The composition of claim 13 , wherein the phospholipid is DSPC.