Respiratory syncytial virus F protein epitopes

Patent metadata

Field	Value
Publication number	US-10016496-B2
Application number	US-201414764635-A
Country	US
Kind code	B2
Filing date	Jan 31, 2014
Priority date	Feb 1, 2013
Publication date	Jul 10, 2018
Grant date	Jul 10, 2018

Abstract

Respiratory Syncytial Virus is a common cold virus belonging to the family of paramyxovirus. RSV is virulent, easily transmissible and the most common cause of lower respiratory tract disease in children of less than 2 years of age. Up to 98% of children attending day care will be infected in a single RSV season. Between 0.5% and 3.2% of children with RSV infection require hospitalization. Approximately 90,000 hospital admissions and 4500 deaths per year were reported in United States. Major risk factors for hospitalization due to RSV are premature birth, chronic lung disease, congenital heart disease, compromised immunity, and age younger than 6 weeks in otherwise healthy children.

First claim

What is claimed is:

1. A fusion protein comprising an isolated Respiratory Syncytial Virus (RSV) fusion (F) protein fragment fused to a heterologous non-RSV F protein polypeptide scaffold;
wherein said non-RSV F protein polypeptide scaffold is a squash family protease inhibitor;
wherein the isolated RSV F protein fragment consists of at least six consecutive amino acids of a peptide consisting of amino acids 195 to 228 of the RSV F protein; and
wherein the inhibitor loop of the squash family protease inhibitor is replaced by the isolated RSV F protein fragment.

2. The fusion protein of claim 1 , wherein said squash family protease inhibitor is the Ecballium elaterium trypsin inhibitor II (EETI-II).

3. A fusion protein comprising an isolated Respiratory Syncytial Virus (RSV) fusion (F) protein fragment fused to a heterologous non-RSV F protein polypeptide scaffold;
wherein said non-RSV F protein polypeptide scaffold is a squash family protease inhibitor;
wherein the isolated RSV F protein fragment consists of a peptide consisting of amino acids 195 to 228 of the RSV F protein; and
wherein the inhibitor loop of the squash family protease inhibitor is replaced by the isolated RSV F protein fragment.

4. The fusion protein of claim 3 , wherein said squash family protease inhibitor is the Ecballium elaterium trypsin inhibitor II (EETI-II).

5. The fusion protein of claim 3 , wherein the cysteine at position 212 of the RSV F protein has been substituted by a serine.

6. The fusion protein of claim 3 , wherein the RSV F protein comprises a human RSV subtype A F protein.

7. The fusion protein of claim 3 , wherein the RSV F protein comprises a human RSV subtype B F protein.

8. The fusion protein of claim 6 , wherein the human RSV subtype A comprises RSV A2.

9. The fusion protein of claim 7 , wherein the human RSV subtype B comprises RSV 18537.

10. An immunogenic composition comprising the fusion protein of claim 3 and a pharmaceutically acceptable carrier.

11. The immunogenic composition of claim 10 further comprising an adjuvant.

12. The fusion protein of claim 1 , wherein the isolated RSV F protein fragment consists of 8 to 10consecutive amino acids of a peptide consisting of amino acids 195 to 228 of the RSV F protein.

13. A method of inducing an immune response to RSV in a subject, the method comprising administering an immunogenic composition of claim 10 to the subject.

14. The fusion protein of claim 1 , wherein the isolated RSV F protein fragment consists of amino acids 195 to 228 of the RSV F protein.

15. The fusion protein of claim 3 , wherein amino acids 195 to 228 of the RSV F protein consists of SEQ ID NO:7 or SEQ ID NO:9.

16. The fusion protein of claim 3 comprising SEQ ID NO:5.

17. A method for identifying an anti-RSV F protein antibody, comprising screening an antibody library for an antibody which binds to the fusion protein of claim 3 , but does not bind to the heterologous non-RSV F protein polypeptide scaffold alone.

18. A fusion protein, the fusion protein comprising the polypeptide scaffold Ecballium elaterium trypsin inhibitor II (EETI-II), wherein the inhibitor loop of EETI-II is replaced by amino acids 1 to 34 of SEQ ID NO:7 or amino acids 1 to 34 of SEQ ID NO:9, and wherein the fusion protein is specifically bound by monoclonal antibody D25 or an antigen-binding fragment thereof.

19. A method for identifying an anti-RSV F protein antibody, comprising screening an antibody library for an antibody which binds to the fusion protein of claim 18 , but does not bind to the heterologous non-RSV F protein polypeptide scaffold alone.

20. The fusion protein of claim 18 , the fusion protein comprising the amino acid sequence SEQ ID NO: 5.

21. An immunogenic composition comprising the fusion protein of claim 18 and a pharmaceutically acceptable carrier and/or adjuvant.

22. A method of inducing an immune response to RSV in a subject, the method comprising administering an immunogenic composition of claim 18 to the subject.

23. The fusion protein of claim 3 , wherein the fusion protein is specifically bound by monoclonal antibody D25 or an antigen-binding fragment thereof.

24. A method for identifying an anti-RSV F protein antibody, comprising screening an antibody library for an antibody which binds to the fusion protein of claim 1 , but does not bind to the heterologous non-RSV F protein polypeptide scaffold alone.

25. An immunogenic composition comprising the fusion protein of claim 1 and a pharmaceutically acceptable carrier and/or adjuvant.

Assignees

• Medimmune Llc

Inventors

• Ulbrandt Nancy

Classifications

• A61P31/14

  for RNA viruses · CPC title

• C07K16/11

  Paramyxoviridae (F); Pneumoviridae (F), e.g. respiratory syncytial virus [RSV] · CPC title

• C07K2317/34

  Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues · CPC title

• C07K2319/00

  Fusion polypeptide · 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