Influenza vaccine

1. A vaccine comprising a messenger ribonucleic acid (mRNA) polynucleotide comprising an open reading frame encoding an influenza virus hemagglutinin antigen formulated in a lipid nanoparticle (LNP), wherein 100% of uracil nucleosides in the open reading frame are N1-methylpseudouridine, wherein one dose of the vaccine contains 10-100 μg of the mRNA polynucleotide, wherein the LNP comprises 20-60 mol % cationic lipid, 5-25 mol % neutral lipid, 25-55 mol % cholesterol, and 0.5-15 mol % PEG-modified lipid, wherein the cationic lipid comprises a cationic lipid of Formula (I):

or a salt or isomer thereof, wherein:

R 1 is —R″M′R′;

R 2 and R 3 are each independently C 1-14 alkyl,

R 4 is —(CH 2 ) n Q where Q is —OR, R is H, and n is 4;

R 5 , R 6 , and R 7 are each H;

M and M′ are each —OC(O)—;

R′ is C 1-18 alkyl;

R″ is C 3-14 alkyl; and

m is 6.

2. The vaccine of claim 1 , wherein the LNP comprises 0.5-5 mol % PEG-modified lipid.

3. The vaccine of claim 2 , wherein the cationic lipid of Formula (I) is Compound 25:

4. The vaccine of claim 1 , wherein the neutral lipid is DSPC, the PEG-modified lipid is PEG-DMG, and the cationic lipid of Formula (I) is Compound 25:

5. The vaccine of claim 4 , wherein the vaccine is formulated for intramuscular injection.

6. A vaccine comprising a first messenger ribonucleic acid (mRNA) polynucleotide comprising an open reading frame encoding a first influenza virus hemagglutinin antigen and a second mRNA polynucleotide comprising an open reading frame encoding a second influenza virus hemagglutinin antigen, wherein 100% of uracil nucleosides in the open reading frames are N1-methylpseudouridine, wherein one dose of the vaccine contains 10-100 μg mRNA polynucleotide, wherein the first and second mRNA polynucleotides are co-formulated in a lipid nanoparticle (LNP) comprising 20-60 mol % cationic lipid, 5-25 mol % neutral lipid, 25-55 mol % cholesterol, and 0.5-15 mol % PEG-modified lipid, wherein the cationic lipid comprises a cationic lipid of Formula (I):

or a salt or isomer thereof, wherein:

R 1 is —R″M′R′;

R 2 and R, are each independently C 1-14 alkyl,

R 4 is —(CH 2 ) n Q where Q is —OR, R is H, and n is 4;

R 5 , R 6 , and R 7 are each H;

M and M′ are each —OC(O)—;

R′ is C 1-18 alkyl;

R″ is C 3-14 alkyl; and

m is 6.

7. The vaccine of claim 6 , wherein the LNP comprises 0.5-5 mol % PEG-modified lipid.

8. The vaccine of claim 7 , wherein the cationic lipid of Formula (I) is Compound 25:

9. The vaccine of claim 6 , wherein the neutral lipid is DSPC, the PEG-modified lipid is PEG-DMG, and the cationic lipid of Formula (I) is Compound 25:

10. The vaccine of claim 9 , wherein the vaccine is formulated for intramuscular injection.

11. The vaccine of claim 6 , further comprising a third mRNA polynucleotide comprising an open reading frame encoding a third influenza virus hemagglutinin antigen, wherein 100% of uracil nucleosides in the open reading frame are N1-methylpseudouridine, wherein the third mRNA polynucleotide is co-formulated in the LNP.

12. The vaccine of claim 11 , wherein the first, second, and third influenza virus hemagglutinin antigens comprise an influenza A H1N1 hemagglutinin antigen, an influenza A H3N2 hemagglutinin antigen, and an influenza B hemagglutinin antigen.

13. A vaccine comprising a first messenger ribonucleic acid (mRNA) polynucleotide comprising an open reading frame encoding a first influenza virus hemagglutinin antigen and a second mRNA polynucleotide comprising an open reading frame encoding a second influenza virus hemagglutinin antigen, wherein 100% of uracil nucleosides in the open reading frames are N1-methylpseudouridine, wherein one dose of the vaccine contains 10-100 μg mRNA polynucleotide, wherein the first and second mRNA polynucleotides are formulated in separate lipid nanoparticles (LNPs), each comprising 20-60 mol % cationic lipid, 5-25 mol % neutral lipid, 25-55 mol % cholesterol, and 0.5-15 mol % PEG-modified lipid, wherein the cationic lipid comprises a cationic lipid of Formula (I):

or a salt or isomer thereof, wherein:

R 1 is —R″M′R′;

R 2 and R 3 are each independently C 1-14 alkyl,

R 4 is —(CH 2 ) n Q where Q is —OR, R is H, and n is 4;

R 5 , R 6 , and R 7 are each H;

M and M′ are each —OC(O)—;

R′ is C 1-18 alkyl;

R″ is C 3-14 alkyl; and

m is 6.

14. The vaccine of claim 13 , wherein the LNPs comprise 0.5-5 mol % PEG-modified lipid.

15. The vaccine of claim 14 , wherein the cationic lipid of Formula (I) is Compound 25:

16. The vaccine of claim 13 , wherein the neutral lipid is DSPC, the PEG-modified lipid is PEG-DMG, and the cationic lipid of Formula (I) is Compound 25:

17. The vaccine of claim 16 , wherein the vaccine is formulated for intramuscular injection.

18. The vaccine of claim 13 , further comprising a third mRNA polynucleotide comprising an open reading frame encoding a third influenza virus hemagglutinin antigen, wherein 100% of uracil nucleosides in the open reading frame are N1-methylpseudouridine, wherein the third mRNA polynucleotide is separately formulated in an LNP comprising 20-60 mol % cationic lipid, 5-25 mol % neutral lipid, 25-55 mol % cholesterol, and 0.5-15 mol % PEG-modified lipid, wherein the cationic lipid comprises a cationic lipid of Formula (I):

or a salt or isomer thereof, wherein:

R 1 is —R″M′R′;

R 2 and R 3 are each independently C 1-14 alkyl,

R 4 is —(CH 2 ) n Q where Q is —OR, R is H, and n is 4;

R 5 , R 6 , and R 7 are each H;

M and M′ are each —OC(O)—;

R′ is C 1-18 alkyl;

R″ is C 3-14 alkyl; and

m is 6.

19. The vaccine of claim 18 , wherein the first, second, and third influenza virus hemagglutinin antigens comprise an influenza A H1N1 hemagglutinin antigen, an influenza A H3N2 hemagglutinin antigen, and an influenza B hemagglutinin antigen.