LIPID NANOPARTICLE MRNA VACCINES

1 . A pharmaceutical composition comprising:

(a) a mRNA comprising a coding sequence encoding a coronavirus spike (S) protein or an antigenic fragment thereof; and

(b) a lipid nanoparticle carrier comprising:

(i) a cationic lipid with the formula III:

or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:

L 1 and L 2 are each independently —O(C═O)—, —(C═O)O—, —C(═O)—, —O—, —S(O) x —, —S—S—, —C(═O)S—, —SC(═O)—, —NR a C(═O)—, —C(═O)NR a —, —NR a C(═O)NR a —, —OC(═O)NR a — or —NR a C(═O)O—;

G 1 and G 2 are each independently unsubstituted C 1 -C 12 alkylene or C 1 -C 12 alkenylene;

G 3 is C 1 -C 24 alkylene, C 1 -C 24 alkenylene, C 3 -C 8 cycloalkylene, or C 3 -C 8 cycloalkenylene;

R a is, at each occurrence, independently H or C 1 -C 12 , alkyl;

R 1 and R 2 are each independently C 6 -C 24 alkyl or C 6 -C 24 alkenyl;

R 3 is OR 5 , CN, —C(═O)OR 4 , —OC(═O)R 4 or —NR 5 C(═O)R 4 ;

R 4 is C 1 -C 12 alkyl;

R 5 is H or C 1 -C 6 alkyl; and

x is 0, 1 or 2; or

(ii) a PEG lipid with the formula (IV)

wherein:

R 8 and R 9 are each independently a straight or branched, saturated or unsaturated alkyl chain containing from 10 to 30 carbon atoms, wherein the alkyl chain is optionally interrupted by one or more ester bonds; and

w has a mean value ranging from 30 to 60; or

(iii) a cationic lipid with the formula I:

or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:

L 1 and L 2 are each independently —O(C═O)—, —(C═O)O— or a carbon-carbon double bond;

R 1a and R 1b are, at each occurrence, independently either (a) H or C 1 -C 12 , alkyl, or (b) R 1a is H or C 1 -C 12 , alkyl, and R 1b together with the carbon atom to which it is bound is taken together with an adjacent R 1b and the carbon atom to which it is bound to form a carbon-carbon double bond;

R 2a and R 2b are, at each occurrence, independently either (a) H or C 1 -C 12 , alkyl, or (b) R 2a is H or C 1 -C 12 , alkyl, and R 2b together with the carbon atom to which it is bound is taken together with an adjacent R 2b and the carbon atom to which it is bound to form a carbon-carbon double bond;

R 1a and R 3b are, at each occurrence, independently either (a) H or C 1 -C 12 , alkyl, or (b) R 1a is H or C 1 -C 12 , alkyl, and R 3b together with the carbon atom to which it is bound is taken together with an adjacent R 3b and the carbon atom to which it is bound to form a carbon-carbon double bond;

R 4a and R 4b are, at each occurrence, independently either (a) H or C 1 -C 12 , alkyl, or (b) R 4a is H or C 1 -C 12 , alkyl, and R 4b together with the carbon atom to which it is bound is taken together with an adjacent R 4b and the carbon atom to which it is bound to form a carbon-carbon double bond;

R 5 and R 6 are each independently methyl or cycloalkyl;

R 7 is, at each occurrence, independently H or C 1 -C 12 , alkyl;

R 8 and R 9 are each independently C 1 -C 12 , alkyl; or R 8 and R 9 , together with the nitrogen atom to which they are attached, form a 5, 6 or 7-membered heterocyclic ring comprising one nitrogen atom;

a and d are each independently an integer from 0 to 24;

b and c are each independently an integer from 1 to 24; and

e is 1 or 2; or

(iv) a cationic liquid with the formula H:

or a pharmaceutically acceptable salt, tautomer or stereoisomer thereof, wherein:

L 1 and L 2 are each independently —O(C═O)—, —(C═O)O—, —C(═O)—, —O—, —S—S—, —C(═O)S—, —SC(═O)—, —NR a C(═O)—, —C(═O)NR a —, —NR a C(═O)NR a —, —OC(═O)NR a —, —NR a C(═O)O—, or a direct bond;

G 1 is C 1 -C 2 alkylene, —(C═O)—, —O(C═O)—, —SC(═O)—, —NR a C(═O)— or a direct bond

G 2 is —C(═O)—, —(C═O)O—, —C(═O)S—, —C(═O)NR a — or a direct bond;

G 3 is C 1 -C 6 alkylene;

R a is, at each occurrence, independently H or C 1 -C 12 alkyl;

R 1a and R 16 are, at each occurrence, independently either: (a) H or C 1 -C 12 alkyl; or (b) R 1a is H or C 1 -C 12 , alkyl, and R 1b together with the carbon atom to which it is bound is taken together with an adjacent R 1b and the carbon atom to which it is bound to form a carbon-carbon double bond;

R 2a and R 2b are, at each occurrence, independently either: (a) H or C 1 -C 12 , alkyl; or (b) R 2a is H or C 1 -C 12 , alkyl, and R 2b together with the carbon atom to which it is bound is taken together with an adjacent R 2b and the carbon atom to which it is bound to form a carbon-carbon double bond;

R 1a and R 3b are, at each occurrence, independently either: (a) H or C 1 -C 12 , alkyl; or (b) R 1a is H or C 1 -C 12 , alkyl, and R 3b together with the carbon atom to which it is bound is taken together with an adjacent R 3b and the carbon atom to which it is bound to form a carbon-carbon double bond;

R 4a and R 4b are, at each occurrence, independently either: (a) H or C 1 -C 12 , alkyl; or (b) R 4a is H or C 1 -C 12 , alkyl, and R 4b together with the carbon atom to which it is bound is taken together with an adjacent R 4b and the carbon atom to which it is bound to form a carbon-carbon double bond;

R 5 and R 6 are each independently H or methyl;

R 7 is C 4 -C 20 alkyl;

R 8 and R 9 are each independently C 1 -C 12 , alkyl; or R 8 and R 9 , together with the nitrogen atom to which they are attached, form a 5, 6 or 7-membered heterocyclic ring;

a, b, c and d are each independently an integer from 1 to 24; and

x is 0, 1 or 2.

2 . The pharmaceutical composition of claim 1 , wherein the mRNA does not comprise a nucleoside having a base modification.

3 . The pharmaceutical composition of claim 1 , wherein the coding sequence of the mRNA consists of A, U, G and C nucleosides.

4 . The pharmaceutical composition of claim 1 , wherein the mRNA comprises at least one chemical modification that is a nucleoside modification

5 . The pharmaceutical composition of claim 1 , wherein the mRNA comprises a 1-methylpseudouridine substitution.

6 . The pharmaceutical composition of claim 1 , wherein the mRNA is encapsulated in or associated with said lipid nanoparticle.

7 . The pharmaceutical composition of claim 1 , wherein the mRNA coding sequence encodes a coronavirus S protein.

8 . The pharmaceutical composition of claim 1 , wherein the mRNA coding sequence encodes an antigenic fragment of a coronavirus S protein.

9 . The pharmaceutical composition of claim 1 , wherein the coronavirus S protein is from a SARS coronavirus.

10 . The pharmaceutical composition of claim 1 , wherein the lipid nanoparticle comprises the cationic lipid of any one of formulae (I), (II), and (III); and additionally comprises:

(a) a PEG lipid with the formula (IV):

wherein:

R 8 and R 9 are each independently a straight or branched, saturated or unsaturated alkyl chain containing from 10 to 30 carbon atoms, wherein the alkyl chain is optionally interrupted by one or more ester bonds; and

w has a mean value ranging from 30 to 60; or

(b) a pegylated diacylglycerol (PEG-DAG).

11 . The pharmaceutical composition of claim 10 , comprising a PEG lipid with the formula (IV):

wherein:

R 8 and R 9 are each independently a straight or branched, saturated or unsaturated alkyl chain containing from 10 to 30 carbon atoms, wherein the alkyl chain is optionally interrupted by one or more ester bonds; and

w has a mean value ranging from 30 to 60.

12 . The pharmaceutical composition of claim 10 , comprising a pegylated diacylglycerol (PEG-DAG).

13 . The pharmaceutical composition of claim 12 , wherein the PEG-DAG is a pegylated 1-(monomethoxy-polyethyleneglycol)-2,3-dimyristoylglycerol (PEG-DMG).

14 . The pharmaceutical composition of claim 1 , wherein the lipid nanoparticle comprises a cationic lipid selected from the structures I-1 to I-41, II-1 to II-34 and III-1 to III-36:

No.

Structure

I-1

I-2

I-3

I-4

I-5

I-6

I-7

I-8

I-9

I-10

I-11

I-12

I-13

I-14

I-15

I-16

I-17

I-18

I-19

I-20

I-21

I-22

I-23

I-24

I-25

I-26

I-27

I-28

I-29

I-30

I-31

I-32

I-33

I-34

I-35

I-36

I-37

I-38

I-39

I-40

I-41

or

No.

Structure

II-1

II-2

II-3

II-4

II-5

II-6

II-7

II-8

II-9

II-10

II-11

II-12

II-13

II-14

II-15

II-16

II-17

II-18

II-19

II-20

II-21

II-22

II-23

II-24

II-25

II-26

II-27

II-28

II-29

II-30

II-31

II-32

II-33

II-34

II-35

II-36

or

No.

Structure

III-1

III-2

III-3

III-4

III-5

III-6

III-7

III-8

III-9

III-10

III-11

III-12

III-13

III-14

III-15

III-16

III-17

III-18

III-19

III-20

III-21

III-22

III-23

III-24

III-25

III-26

III-27

III-28

III-29

III-30

III-31

III-32

III-33

III-34

III-35

III-36

15 . The composition of claim 1 , wherein in the PEG lipid R 8 and R 9 are saturated alkyl chains.

16 . The lipid nanoparticle according to claim 14 , wherein the PEG lipid is

wherein n is an integer selected such that the average molecular weight of the PEG lipid is about 2500 g/mol.

17 . The pharmaceutical composition of claim 1 , wherein the lipid nanoparticle comprises the cationic lipid of formula (I), (II) or (III), DSPC, cholesterol and a PEG-lipid.

18 . The pharmaceutical composition of claim 1 , wherein the mRNA sequence additionally comprises:

a) a 5′-CAP structure;

b) a poly(A) sequence;

c) a poly (C) sequence; or

d) two or more of a), b) and c).

19 . The pharmaceutical composition of claim 1 , wherein the mRNA sequence additionally comprises at least one histone stem loop.

20 . The pharmaceutical composition of claim 19 , wherein the mRNA sequence comprises, in 5′ to 3′-direction, the following elements:

a) a 5′ m7GpppN CAP structure,

b) a coding sequence encoding a coronavirus S protein or an antigenic fragment thereof,

c) a poly(A) sequence of 10 to 200 adenosine nucleotides,

d) optionally a poly(C) sequence, and

e) optionally a histone stem-loop.

21 . The pharmaceutical composition of claim 20 , wherein the mRNA sequence comprises, in 5′ to 3′-direction, the following elements:

a) a 5′ CAP1 structure,

b) a coding sequence encoding a coronavirus S protein or an antigenic fragment thereof,

c) a 3′-UTR element comprising a sequence from an alpha globin gene;

d) a poly(A) sequence of 10 to 200 adenosine nucleotides,

e) a poly(C) sequence of 10 to 200 cytosine nucleotides, and

f) a histone stem-loop.

22 . The pharmaceutical composition of claim 1 , wherein the mRNA sequence comprises, in 5′ to 3′-direction, the following elements:

a) a 5′ CAP1 structure,

b) a 5′-UTR element which comprises a nucleic acid sequence from the 5′-UTR of a TOP gene;

c) a coding sequence encoding a coronavirus S protein or an antigenic fragment thereof,

d) a 3′-UTR sequence;

e) optionally, a histone stem-loop; and

f) a poly(A) sequence of 10 to 200 adenosine nucleotides.

23 . The pharmaceutical composition of claim 22 , wherein (b) the 5′-UTR sequence is a sequence from a HSD17B4 gene 5′-UTR.

24 . The pharmaceutical composition of claim 1 , wherein the lipid nanoparticle comprises the cationic lipid of formula (I).

25 . The pharmaceutical composition of claim 1 , wherein the lipid nanoparticle comprises the cationic lipid of formula (II).

26 . The pharmaceutical composition of claim 1 , wherein the lipid nanoparticle comprises the cationic lipid of formula (III).

27 . A method of treating or preventing a disease or disorder in a subject in need thereof comprising administering to the subject an effective amount of a pharmaceutical composition of claim 1 .

28 . A method for raising an immune response in a subject in need thereof, comprising administering to the subject an effective amount of a pharmaceutical composition of claim 1 .