Prefusion RSV F proteins and their use

Patent metadata

Field	Value
Publication number	US-10858400-B2
Application number	US-201715633578-A
Country	US
Kind code	B2
Filing date	Jun 26, 2017
Priority date	Mar 13, 2013
Publication date	Dec 8, 2020
Grant date	Dec 8, 2020

Abstract

Disclosed are immunogens including a recombinant RSV F protein stabilized in a prefusion conformation. Also disclosed are nucleic acids encoding the immunogens and methods of producing the immunogens. Methods for generating an immune response in a subject are also disclosed. In some embodiments, the method is a method for treating or preventing a RSV infection in a subject by administering a therapeutically effective amount of the immunogen to the subject.

First claim

We claim:

1. An isolated immunogen, comprising:
a recombinant RSV F protein comprising S 190F and V207L amino acid substitutions compared to a native RSV F protein that stabilize the recombinant RSV F protein in a prefusion conformation;
wherein the prefusion conformation comprises an antigenic site Ø comprising RSV F residues 62-69 and 196-209 that specifically binds to a D25 antibody or an AM22 antibody after incubation at 20° C. in phosphate buffered saline at physiological pH for at least 24 hours in the absence of the D25 antibody or the AM22 antibody.

2. The immunogen of claim 1 , wherein the RSV F protein further comprises one or more additional amino acid substitutions.

3. The immunogen of claim 1 , wherein the RSV F protein is a human subtype A or subtype B, or bovine RSV F protein comprising the amino acid substitutions.

4. The immunogen of claim 1 , wherein the recombinant RSV F protein comprises a F 2 polypeptide and a F 1 polypeptide comprising or consisting of RSV F positions 26-109 and 137-513, respectively.

5. The immunogen of claim 1 , wherein the recombinant RSV F protein comprises a F2 polypeptide and a F 1 polypeptide, wherein a C-terminal residue of the F 2 polypeptide is linked to an N-terminal residue of the F 1 polypeptide by a heterologous peptide linker.

6. The immunogen of claim 1 , wherein the recombinant RSV F protein comprises an F 1 polypeptide comprising the amino acid sequence set forth as residues 137-513 of SEQ ID NO: 191.

7. The immunogen of claim 1 , wherein the recombinant RSV F protein is soluble and comprises an F 1 ectodomain comprising a C-terminal residue linked to a trimerization domain.

8. The immunogen of claim 7 , wherein the trimerization domain is a Foldon domain.

9. The immunogen of claim 8 , wherein the recombinant RSV F protein linked to the trimerization domain comprises the amino acid sequences set forth as positions 26-109 and 137-544 of SEQ ID NO: 191.

10. The immunogen of claim 1 , wherein the C-terminus of recombinant RSV F protein is linked to a ferritin domain, an encapsulin domain, a Sulfur Oxygenase Reductase (SOR) domain, a lumazine synthase domain, or a pyruvate dehydrogenase domain.

11. The immunogen of claim 1 , wherein the recombinant RSV F protein comprises a F 1 ectodomain comprising a C-terminal residue linked to a transmembrane domain.

12. A virus-like particle comprising the immunogen of claim 1 .

13. A protein nanoparticle comprising the immunogen of claim 1 .

14. The protein nanoparticle of claim 13 , wherein the protein nanoparticle is a ferritin nanoparticle, an encapsulin nanoparticle, a Sulfur Oxygenase Reductase (SOR) nanoparticle, a lumazine synthase nanoparticle, or a pyruvate dehydrogenase nanoparticle.

15. A nucleic acid molecule encoding the immunogen of claim 1 .

16. A vector comprising the nucleic acid molecule of claim 15 .

17. An isolated host cell comprising the vector of claim 16 .

18. An immunogenic composition comprising an effective amount of the immunogen of claim 1 , and a pharmaceutically acceptable carrier.

19. A method for generating an immune response to RSV F protein in a subject, comprising administering an effective amount of the immunogen of claim 1 to the subject to generate the immune response.

Assignees

• Us Health

Inventors

• Kwong Peter
• Graham Barney
• Mclellan Jason
• Chen Man
• Zhang Baoshan
• Zhou Tongqing
• Joyce Michael Gordon
• Yang Yongping

Classifications

• C07K2319/70

  containing domain for protein-protein interaction · CPC title

• C07K14/005 Primary

  from viruses · CPC title

• C07K2319/21

  containing a His-tag · CPC title

• C07K2319/50

  containing protease site · CPC title

• C12N2760/18534

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title