Prefusion PIV F immunogens and their use

Patent metadata

Field	Value
Publication number	US-11845778-B2
Application number	US-202117378587-A
Country	US
Kind code	B2
Filing date	Jul 16, 2021
Priority date	Oct 25, 2016
Publication date	Dec 19, 2023
Grant date	Dec 19, 2023

Abstract

Embodiments of a recombinant human Parainfluenza Virus (hPIV) F ectodomain trimer stabilized in a prefusion conformation are provided. Also disclosed are nucleic acids encoding the hPIV F ectodomain trimer and methods of producing the hPIV 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 inhibiting a hPIV infection in a subject by administering a effective amount of the recombinant hPIV F ectodomain trimer to the subject.

First claim

The invention claimed is:

1. An immunogen, comprising:
a recombinant human parainfluenza virus 1 (hPIV1) F ectodomain trimer comprising protomers comprising one or more amino acid substitutions that stabilize the hPIV1 ectodomain trimer in a prefusion conformation, wherein the one or more amino acid substitutions comprise A466I and S473I cavity filling substitutions, wherein the amino acid numbering is according to a reference hPIV1 F sequence set forth as SEQ ID NO: 2.

2. The immunogen of claim 1 , wherein the protomers further comprise a mutation to inhibit cleavage of a F 2 /F 1 protease cleavage site.

3. The immunogen of claim 2 , wherein the mutation comprises a deletion of hPIV1 residues 113 and 114 with residues 112 and 115 linked by a heterologous peptide linker.

4. The immunogen of claim 3 , wherein the heterologous peptide linker consists of the amino acid sequence set forth as GS.

5. The immunogen of claim 1 , wherein one or more amino acid substitutions further comprise a non-native disulfide bond between 175C-241C, 173C-245C, 206C-233C, 88C-225C, 219C-224C, or 165C-171C substitutions.

6. The immunogen of claim 1 , wherein a C-terminal residue of the protomers is one of hPIV1 F residues 473-497.

7. The immunogen of claim 1 , wherein the protomers comprise or consist essentially of hPIV1 residues 22-112 and 115-479 with residues 112 and 115 linked by a heterologous peptide linker and comprise the one or more amino acid substitutions to stabilize the trimer in the prefusion conformation.

8. The immunogen of claim 1 , wherein the protomers comprise or consist essentially of an amino acid sequence at least 90% identical to residues 1-458 of SEQ ID NO: 4, wherein the protomers comprise the one or more amino acid substitutions.

9. The immunogen of claim 8 , wherein the protomers comprise or consist essentially of the amino acid sequence set forth as residues 1-458 of SEQ ID NO: 4.

10. The immunogen of claim 1 , wherein a C-terminal residue of the protomers in the ectodomain is linked to a trimerization domain by a peptide linker, or is directly linked to the trimerization domain.

11. The immunogen of claim 10 , wherein the trimerization domain is a GCN4 trimerization domain.

12. The immunogen of claim 10 , wherein the protomers linked to the trimerization domain comprise or consist essentially of an amino acid sequence at least 90% identical to residues 1-458 of SEQ ID NO: 4, wherein the protomers comprise the one or more amino acid substitutions.

13. The immunogen of claim 12 , wherein the protomers linked to the trimerization domain comprise or consist essentially of the amino acid sequence set forth as SEQ ID NO: 4.

14. The immunogen of claim 1 , wherein the protomers further comprise one or more additional amino acid substitutions.

15. The immunogen of claim 1 , wherein the recombinant hPIV1 F ectodomain trimer is soluble.

16. The immunogen of claim 1 , wherein the C-terminal residue of the protomers in the F ectodomain is linked to a transmembrane domain by a peptide linker, or is directly linked to the transmembrane domain.

17. The immunogen of claim 1 , wherein the C-terminal residue of the protomers in the F ectodomain is linked to a protein nanoparticle subunit by a peptide linker, or is directly linked to the protein nanoparticle subunit.

18. A protein nanoparticle, comprising the immunogen of claim 1 .

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

20. An isolated nucleic acid molecule encoding a protomer of the recombinant hPIV1 F ectodomain trimer of claim 1 .

21. An immunogenic composition comprising the immunogen of claim 1 , a virus like particle comprising the immunogen, a protein nanoparticle comprising the immunogen, or a nucleic acid molecule encoding the immunogen.

22. A method of inducing an immune response to hPIV1 F protein in a subject, comprising administering to the subject an effective amount of the immunogen of claim 1 , a virus like particle comprising the immunogen, a protein nanoparticle comprising the immunogen, or a nucleic acid molecule encoding the immunogen to generate the immune response.

23. The method of claim 22 , wherein the immune response inhibits an hPIV1 infection.

Assignees

• Us Health
• Inst Res Biomedicine

Inventors

• Zhang Baoshan
• Stewart-Jones Guillaume
• Mascola John
• Xu Kai
• Chuang Gwo-Yu
• Ou Li
• Kwong Peter
• Tsybovsky Yaroslav
• Kong Wing-Pui
• Druz Aliaksandr
• Corti Davide
• Lanzavecchia Antonio

Classifications

• C07K14/005 Primary

  from viruses · CPC title

• A61K39/155

  Paramyxoviridae, e.g. parainfluenza virus · CPC title

• A61P31/14

  for RNA viruses · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• C07K2319/50

  containing protease site · CPC title