Prefusion RSV F proteins and their use

Patent metadata

Field	Value
Publication number	US-12275758-B2
Application number	US-202418409645-A
Country	US
Kind code	B2
Filing date	Jan 10, 2024
Priority date	Mar 29, 2016
Publication date	Apr 15, 2025
Grant date	Apr 15, 2025

Abstract

Embodiments of a recombinant Respiratory Syncytial Virus (RSV) F ectodomain trimer stabilized in a prefusion conformation are provided. Also disclosed are nucleic acids encoding the RSV F ectodomain trimer and methods of producing the RSV F ectodomain trimer. Methods for inducing an immune response in a subject are also disclosed. In some embodiments, the method can be a method for treating or preventing a RSV infection in a subject by administering a therapeutically effective amount of the recombinant RSV F ectodomain trimer to the subject.

First claim

We claim:

1. A ribonucleic acid (RNA) molecule encoding a recombinant respiratory syncytial virus (RSV) F protein comprising:
a deletion of RSV F positions 104-144 and a glycine-serine peptide linker between RSV F positions 103 and 145,
S155C and S290C amino acid substitutions, S190F and V207L amino acid substitutions, and A149C and Y458C amino acid substitutions;
wherein the amino acid deletion and substitutions are according to the reference RSV sequence set forth as SEQ ID NO: 57; and
wherein the recombinant RSV F protein comprises an amino acid sequence set forth as residues 26-474 of any one of SEQ ID NOs: 1, 5, or 10, or residues 31-479 of any one of SEQ ID NOs: 15 or 20.

2. The RNA molecule of claim 1 , wherein the RSV F protein comprises the amino acid sequence set forth as residues 26-474 of SEQ ID NO: 1.

3. The RNA molecule of claim 1 , wherein the RSV F protein comprises the amino acid sequence set forth as residues 26-474 of SEQ ID NO: 5.

4. The RNA molecule of claim 1 , wherein the RSV F protein comprises the amino acid sequence set forth as residues 26-474 of SEQ ID NO: 10.

5. The RNA molecule of claim 1 , wherein the RSV F protein comprises the amino acid sequence set forth as residues 31-479 of SEQ ID NO: 15.

6. The RNA molecule of claim 1 , wherein the RSV F protein comprises the amino acid sequence set forth as residues 31-479 of SEQ ID NO: 20.

7. The RNA molecule of claim 1 , wherein the RSV F protein comprises an RSV F transmembrane domain and cytoplasmic tail.

8. The RNA molecule of claim 1 , wherein the RNA molecule encodes a precursor protein of the RSV F protein comprising, from N to C terminal, a signal peptide, a F 2 polypeptide, an F 1 ectodomain, and a transmembrane domain and cytosolic tail.

9. An immunogenic composition comprising the RNA molecule of claim 1 .

10. A method of inducing an immune response to RSV F protein in a subject, comprising administering to the subject an effective amount of the RNA molecule of claim 1 to generate the immune response.

11. The method of claim 10 , wherein the subject is a human or a bovine subject.

12. The method of claim 10 , wherein the subject is in the third trimester of pregnancy.

13. The method of claim 10 , wherein the subject is less than one year old.

14. The method of claim 10 , wherein the immune response inhibits RSV infection in the subject.

15. A messenger ribonucleic acid (mRNA) molecule encoding a recombinant respiratory syncytial virus (RSV) F protein comprising:
a deletion of RSV F positions 104-144 and a glycine-serine peptide linker between RSV F positions 103 and 145,
S155C and S290C amino acid substitutions, S190F and V207L amino acid substitutions, and A149C and Y458C amino acid substitutions;
wherein the amino acid deletion and substitutions are according to the reference RSV sequence set forth as SEQ ID NO: 57; and
wherein the recombinant RSV F protein comprises an amino acid sequence set forth as residues 26-474 of SEQ ID NO: 1, 5, or 10, or residues 31-479 of SEQ ID NO: 15 or 20.

16. The mRNA molecule of claim 15 , wherein the RSV F protein comprises the amino acid sequence set forth as residues 26-474 of SEQ ID NO: 1.

17. The mRNA molecule of claim 15 , wherein the RSV F protein comprises the amino acid sequence set forth as residues 26-474 of SEQ ID NO: 5.

18. The mRNA molecule of claim 15 , wherein the RSV F protein comprises the amino acid sequence set forth as residues 26-474 of SEQ ID NO: 10.

19. The mRNA molecule of claim 15 , wherein the RSV F protein comprises the amino acid sequence set forth as residues 31-479 of SEQ ID NO: 15.

20. The mRNA molecule of claim 15 , wherein the RSV F protein comprises the amino acid sequence set forth as residues 31-479 of SEQ ID NO: 20.

21. The mRNA molecule of claim 15 , wherein the RSV F protein comprises an RSV F transmembrane domain and cytoplasmic tail.

22. The mRNA molecule of claim 15 , wherein the mRNA molecule encodes a precursor protein of the RSV F protein comprising, from N to C terminal, a signal peptide, a F 2 polypeptide, an F 1 ectodomain, and a transmembrane domain and cytosolic tail.

23. An immunogenic composition comprising the mRNA molecule of claim 15 .

24. A method of inducing an immune response to RSV F protein in a subject, comprising administering to the subject an effective amount of the mRNA molecule of claim 15 to generate the immune response.

25. The method of claim 24 , wherein the subject is a human or a bovine subject.

26. The method of claim 24 , wherein the immune response inhibits RSV infection in the subject.

27. The method of claim 24 , wherein the subject is in the third trimester of pregnancy.

28. The method of claim 24 , wherein the subject is less than one year old.

29. A ribonucleic acid (RNA) molecule encoding a recombinant respiratory syncytial virus (RSV) F protein comprising:
a deletion of RSV F positions 104-144 and a glycine-serine peptide linker between RSV F positions 103 and 145,
S155C and S290C amino acid substitutions and S190F and V207L amino acid substitutions; and
(A) Q98C and Q361C substitutions; or
(B) N183GC and N428C substitutions, and an amino acid sequence set forth as residues 26-475 of SEQ ID NO: 2, 6, 11, 30-35, 38-39, 42-43, 46-47, 50-51, or 54-55, or residues 31-480 of SEQ ID NO: 16 or 21; and
wherein the amino acid deletion and substitutions are according to a reference RSV F sequence set forth as SEQ ID NO: 57.

Assignees

• Us Health

Inventors

• Kwong Peter
• Graham Barney
• Mascola John
• Ou Li
• Druz Aliaksandr
• Chen Man
• Kong Wing-Pui
• Georgiev Ivelin Stefanov
• Rundlet Emily
• Joyce Michael Gordon
• Tsybovsky Yaroslav
• Thomas Paul
• Pancera Marie
• Sastry Mallika
• Soto Cinque
• Van Galen Joseph
• Stewart-Jones Guillaume
• Yang Yongping
• Zhang Baoshan
• Baxa Ulrich

Classifications

• C12N2760/18523

  Virus like particles [VLP] · CPC title

• A61K39/00

  Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53) · CPC title

• A61P31/14

  for RNA viruses · CPC title

• A61P37/04

  Immunostimulants · CPC title

• C07K2319/22

  containing a Strep-tag · CPC title