Stabilized RSV F proteins and uses thereof

1. A recombinant respiratory syncytial virus (RSV) F protein trimer, comprising: an amino acid sequence set forth in any one of SEQ ID NOS: 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 44, or 45.

2. An RSV immunogenic composition comprising the recombinant RSV F protein trimer of claim 1 and a pharmaceutically acceptable excipient.

3. The immunogenic composition of claim 2 formulated in an effective amount to produce an antigen-specific immune response directed against the recombinant RSV F trimer or the recombinant RSV F peptide.

4. The immunogenic composition of claim 2 that is formulated with an adjuvant.

5. The immunogenic composition of claim 4 , wherein the adjuvant is an aluminum adjuvant.

6. The immunogenic composition of claim 5 , wherein the aluminum adjuvant is amorphous aluminum hydroxyphosphate sulfate or an aqueous suspension of aluminum hydroxyphosphate.

7. The immunogenic composition of claim 2 further comprising a lipid nanoparticle comprising one or more cationic lipids and a poly(ethyleneglycol)-lipid (PEG lipid).

8. The immunogenic composition of claim 7 , wherein the one or more cationic lipids are ionizable cationic lipids.

9. The immunogenic composition of claim 8 , wherein the ionizable cationic lipids are selected from DLinDMA; DlinKC2DMA; DLin-MC3-DMA; CLinDMA; S-Octyl CLinDMA;

(2 S)-1-{7-[(3 β)-cholest-5-en-3-yloxy]heptyloxy}-3-[(4 Z)-dec-4-en-1-yloxy]-N, N-dimethylpropan-2-amine;

(2 R)-1-{4-[(3β)-cholest-5-en-3-yloxy]butoxy}-3-[(4 Z)-dec-4-en-1-yloxy]-N, N-dimethylpropan-2-amine;

1-[(2 R)-1-{7-[(3 β)-cholest-5-en-3-yloxy]butoxy}-3-(octyloxy)propan-2-yl]guanidine;

1-[(2 R)-1-{7-[(3 β)-cholest-5-en-3-yloxy]heptyloxy}-N,N-dimethyl-3-[(9 Z, 12 Z)-octadeca-9,12-dien-1-yl]oxy]propan-2-amine;

1-[(2 R)-1-{4-[(3 β)-cholest-5-en-3-yl]oxy]butoxy}-N,N-dimethyl-3-[(9 Z, 12 Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine;

(2S)-1-({6-[(3 β)-cholest-5-en-3-yloxy]hexyl}oxy)-N,N-dimethyl-3-[(9 Z)-octadec-9-en-1-yloxy]propan-2-amine;

(3β)-3-[6-{[(2S)-3-[(9Z)-octadec-9-en-1-yloxyl]-2-(pyrrolidin-1-yl)propyl]oxy}hexyl)oxy]cholest-5-ene;

(2R)-1-({4-[(3β)-cholest-5-en-3-yloxy]butoxy}-3-(octyloxy)propan-2-amine;

(2R)-1-({8-[(3β)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-(pentyloxy)propan-2-amine;

(2R)-1-({8-[(3β)-cholest-5-en-3-yloxyl]octyl}oxy)-3-(heptyloxy)-N,N-dimethylpropan-2-amine;

(2R)-1-({8-[(3β)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(2Z)-pent-2-en-1-yloxy]propan-2-amine;

(2S)-1-butoxy-3-({8-[(3β)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethylpropan-2-amine;

(2S-1-({8-[(3β)-cholest-5-en-3-yloxy]octyl}oxy)-[2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9-hexadecafluorononyl)oxy]-N,N-dimethylpropan-2-amine;

2-amino-2-{[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]methyl}propane-1,3-diol;

2-amino-3-((9-(((3S,10R,13R)-10,13-dimethyl-17-(6-methylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)nonyl)oxy)-2-((((9Z,12Z)-octadeca-9,12-dien-1-yl)oxy)methyl)propan-1-ol;

2-amino-3-((6-(((3 S,10R,13R)-10,13-dimethyl-17-(6-methylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)hexyl)oxy)-2-((((Z)-octadec-9-en-1-yl)oxy)methyl)propan-1-ol;

(20Z,23Z)-N,N-dimethylnonacosa-20,23-dien-10-amine;

(17Z,20Z)-N,N-dimethylhexacosa-17,20-dien-9-amine;

(16Z,19Z)-N,N-dimethylpentacosa-16,19-dien-8-amine;

(13Z,16Z)-N,N-dimethyldocosa-13,16-dien-5-amine;

(12Z,15Z)-N,N-dimethylhenicosa-12,15-dien-4-amine;

(14Z,17Z)-N,N-dimethyltricosa-14,17-dien-6-amine;

(15Z,18Z)-N,N-dimethyltetracosa-15,18-dien-7-amine;

(18Z,21Z)-N,N-dimethylheptacosa-18,21-dien-10-amine;

(15Z,18Z)-N,N-dimethyltetracosa-15,18-dien-5-amine;

(14Z,17Z)-N,N-dimethyltricosa-14,17-dien-4-amine;

(19Z,22Z)-N,N-dimethyloctacosa-19,22-dien-9-amine;

(18Z,21Z)-N,N-dimethylheptacosa-18,21-dien-8-amine;

(17Z,20Z)-N,N-dimethylhexacosa-17,20-dien-7-amine;

(16Z,19Z)-N,N-dimethylpentacosa-16,19-dien-6-amine;

(22Z,25Z)-N,N-dimethylhentriaconta-22,25-dien-10-amine;

(21Z,24Z)-N,N-dimethyltriaconta-21,24-dien-9-amine;

(18Z)-N,N-dimethylheptacos-18-en-10-amine;

(17Z)-N,N-dimethylhexacos-17-en-9-amine;

(19Z,22Z)-N,N-dimethyloctacosa-19,22-dien-7-amine;

N,N-dimethylheptacosan-10-amine;

(20Z,23Z)-N-ethyl-N-methylnonacosa-20,23-dien-10-amine;

1-[(11Z,14Z)-1-nonylicosa-11,14-dien-1-yl]pyrrolidine;

(20Z)-N,N-dimethylheptacos-20-en-10-amine;

(15Z)-N,N-dimethylheptacos-15-en-10-amine;

(14Z)-N,N-dimethylnonacos-14-en-10-amine;

(17Z)-N,N-dimethylnonacos-17-en-10-amine;

(24Z)-N,N-dimethyltritriacont-24-en-10-amine;

(20Z)-N,N-dimethylnonacos-20-en-10-amine;

(22Z)-N,N-dimethylhentriacont-22-en-10-amine;

(16Z)-N,N-dimethylpentacos-16-en-8-amine;

(12Z,15Z)-N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine;

(13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine;

N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]heptadecan-8-amine;

1-[(1S,2R)-2-hexylcyclopropyl]-N,N-dimethylnonadecan-10-amine;

N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]nonadecan-10-amine;

N,N-dimethyl-21-[(1S,2R)-2-octylcyclopropyl]henicosan-10-amine;

N,N-dimethyl-1-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]methyl}cyclopropyl]nonadecan-10-amine;

N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]hexadecan-8-amine;

N,N-dimethyl-1-[(1R,2S)-2-undecylcyclopropyl]tetradecan-5-amine;

N,N-dimethyl-3- {7- [(1 S,2R)-2-octylcyclopropyl]heptyl}dodecan-1-amine;

1-[(1R,2S)-2-heptylcyclopropyl]-N,N-dimethyloctadecan-9-amine;

1-[(1S,2R)-2-decylcyclopropyl]-N,N-dimethylpentadecan-6-amine;

N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]pentadecan-8-amine; and

(11E,20Z,23Z)-N,N-dimethylnonacosa-11,20,23-trien-10-amine;

or a pharmaceutically acceptable salt thereof, or a stereoisomer of any of the foregoing.

10. The immunogenic composition of claim 7 , further comprising one or more non-cationic lipids selected from a phospholipid, a phospholipid derivative, a fatty acid, sterol, or a combination thereof.

11. The immunogenic composition of claim 10 , which comprises 34-59 mole % ionizable cationic lipids selected from the group consisting of (2S)-1-({6-[(3β))-cholest-5-en-3-yloxy]hexyl}oxy)-N,N-dimethyl-3-[(9 Z)-octadec-9-en-1-yloxy]propan-2-amine; (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine; and N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]heptadecan-8-amine, 30-48 mole % cholesterol, 10-24% DSPC and 1-2 mole % PEG-DMG.

12. A method of producing an immune response to RSV F protein in a subject, the method comprising administering to the subject an effective amount of the RSV F protein trimer of claim 1 .

13. The method of claim 12 , wherein the subject is administered a single dose of the RSV F protein trimer.

14. The method of claim 12 , wherein the subject is administered at least one booster dose of the RSV F protein trimer.

15. The method of claim 12 , wherein the RSV F protein trimer is administered to the subject by intradermal injection or intramuscular injection.

16. The method of claim 12 , wherein the subject has been exposed to RSV, wherein the subject is infected with RSV, or wherein the subject is at risk of infection by RSV.