MRNA-loaded lipid nanoparticles and processes of making the same

1 . A lipid nanoparticle for the delivery of mRNA to a subject in need thereof, the lipid nanoparticle comprising

one or more cationic lipids at a concentration between 30 molar percent and 60 molar percent,

one or more PEG-modified lipids at a concentration between 1 molar percent and 15 molar percent,

one or more helper lipids comprising 1,2-Dierucoyl-sn-glycero-3-phosphoethanolamine (DEPE), and wherein the DEPE is present at a concentration between 25 molar percent and 35 molar percent, and

one or more sterols at a concentration between 20 molar percent and 30 molar percent,

wherein the molar percents are of total lipids in the lipid nanoparticle and wherein the lipid nanoparticle encapsulates the mRNA.

2 . The lipid nanoparticle of claim 1 , wherein the one or more cationic lipids are or comprise a cationic lipid that comprises one to four alkyl chains, each of C 10 -C 16 in length.

3 . The lipid nanoparticle of claim 1 , wherein the one or more cationic lipids are or comprise a cationic lipid of the following formula:

or a pharmaceutically acceptable salt thereof; wherein

each of R 1 and R 2 is independently H or C 1 -C 6 aliphatic;

each m is independently an integer having a value of 1 to 4;

each A is independently a covalent bond or arylene;

each L 1 is independently an ester, thioester, disulfide, or anhydride group;

each L 2 is independently C 2 -C 10 aliphatic;

each X 1 is independently H or OH; and

each R 3 is independently C 6 -C 20 aliphatic.

4 . The lipid nanoparticle of claim 1 , wherein the one or more PEG-modified lipids comprise a poly (ethylene) glycol chain of up to 5 kDa covalently attached to a lipid with alkyl chain(s) of C 6 -C 20 in length.

5 . The lipid nanoparticle of claim 1 , wherein the mRNA encodes a therapeutic protein or peptide.

6 . A pharmaceutical composition comprising a plurality of the lipid nanoparticle of claim 1 , wherein greater than 70% of the lipid nanoparticles in the composition have a size ranging from 75 nm to 150 nm.

7 . The lipid nanoparticle of claim 1 , wherein the one or more cationic lipids are a lipidoid comprising four aliphatic chains.

8 . The lipid nanoparticle of claim 1 , wherein the mRNA encodes an antigen.

9 . The lipid nanoparticle of claim 1 , wherein the mRNA encodes an antibody light chain or an antibody heavy chain.

10 . The lipid nanoparticle of claim 1 , wherein the mRNA encodes a polypeptide absent or deficient in the subject.

11 . The lipid nanoparticle of claim 3 , wherein each R 3 is independently C 8 -C 16 aliphatic.

12 . The lipid nanoparticle of claim 1 , wherein the one or more sterols comprise a cholesterol or PEGylated cholesterol, or a combination thereof.

13 . The lipid nanoparticle of claim 1 , wherein

the one or more cationic lipids are present at a concentration of about 40 molar percent,

the one or more PEG-modified lipids are present at a concentration of about 3 molar percent,

the DEPE is present at a concentration of about 32 molar percent, and

the one or more sterols are present at a concentration of about 25 molar percent,

wherein the molar percents are of total lipids in the lipid nanoparticle.

14 . The lipid nanoparticle of claim 13 ,

wherein the one or more sterols comprise a cholesterol.

15 . The lipid nanoparticle of claim 1 , wherein the one or more cationic lipids are or comprise Compound 1:

or a pharmaceutically acceptable salt thereof.

16 . The lipid nanoparticle of claim 1 , wherein the one or more cationic lipids are or comprise cKK-E12 or a pharmaceutically acceptable salt thereof.

17 . The lipid nanoparticle of claim 1 , wherein the one or more cationic lipids are or comprise imidazole cholesterol ester or a pharmaceutically acceptable salt thereof.

18 . The lipid nanoparticle of claim 1 , wherein the one or more PEG-modified lipids are or comprise 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000.