Lipid nanoparticle compositions and methods of formulating the same
Provided herein are compositions and methods of reducing adduct formation.
1. A composition comprising a lipid nanoparticle comprising a mRNA, a phospholipid, a cholesterol, a PEG-lipid, and an ionizable lipid,
wherein the mRNA comprises an N1-methyl-pseudouridine (m1ψ),
wherein the ionizable lipid is selected from
wherein the composition comprises a Tris (tris(hydroxymethyl)aminomethane) buffer and sucrose, and
wherein less than about 10% of the mRNA is in the form of an ionizable lipid-polynucleotide adduct impurity, as measured by reverse phase ion pair high performance liquid chromatography (RP-IP HPLC).
2. The composition of claim 1 , wherein the ionizable lipid is
3. The composition of claim 2 , wherein less than about 5% of the mRNA is in the form of the ionizable lipid-polynucleotide adduct impurity.
4. The composition of claim 2 , wherein less than about 1% of the mRNA is in the form of the ionizable lipid-polynucleotide adduct impurity.
5. The composition of claim 2 , wherein the composition comprises a molar ratio of 20-60% ionizable lipid, 5-25% phospholipid, 25-55% cholesterol, and 0.5-15% PEG-lipid, based on the lipid components.
6. The composition of claim 2 , wherein all uridines in the mRNA are N1-methyl-pseudouridines (m1ψ).
7. The composition of claim 2 , wherein the ionizable lipid-polynucleotide adduct impurity comprises an aldehyde-mRNA adduct impurity.
8. The composition of claim 7 , wherein an amount of lipid aldehydes in the composition is less than about 50 ppm.
9. The composition of claim 7 , wherein an amount of the ionizable lipid-polynucleotide adduct impurity increases at an average rate of less than about 2% per day when stored at a temperature of about 25° C. or below.
10. The composition of claim 7 , wherein an amount of the ionizable lipid-polynucleotide adduct impurity increases at an average rate of less than about 0.5% per day when stored at a temperature of about 5° C. or below.
11. The composition of claim 7 , wherein an amount of the ionizable lipid-polynucleotide adduct impurity increases at an average rate of less than about 0.5% per day when stored at a refrigerated temperature.
12. The composition of claim 11 , wherein the refrigerated temperature is about 5° C.
13. The composition of claim 1 , wherein the ionizable lipid is
14. The composition of claim 13 , wherein less than about 5% of the mRNA is in the form of the ionizable lipid-polynucleotide adduct impurity.
15. The composition of claim 13 , wherein less than about 1% of the mRNA is in the form of the ionizable lipid-polynucleotide adduct impurity.
16. The composition of claim 13 , wherein the composition comprises a molar ratio of 20-60% ionizable lipid, 5-25% phospholipid, 25-55% cholesterol, and 0.5-15% PEG-lipid, based on the lipid components.
17. The composition of claim 13 , wherein all uridines in the mRNA are N1-methyl-pseudouridines (m1ψ).
18. The composition of claim 13 , wherein the ionizable lipid-polynucleotide adduct impurity comprises an aldehyde-mRNA adduct impurity.
19. The composition of claim 18 , wherein an amount of lipid aldehydes in the composition is less than about 50 ppm.
20. The composition of claim 18 , wherein an amount of the ionizable lipid-polynucleotide adduct impurity increases at an average rate of less than about 2% per day when stored at a temperature of about 25° C. or below.
21. The composition of claim 18 , wherein an amount of the ionizable lipid-polynucleotide adduct impurity increases at an average rate of less than about 0.5% per day when stored at a temperature of about 5° C. or below.
22. The composition of claim 18 , wherein an amount of the ionizable lipid-polynucleotide adduct impurity increases at an average rate of less than about 0.5% per day when stored at a refrigerated temperature.
23. The composition of claim 22 , wherein the refrigerated temperature is about 5° C.