STABILIZED SOLUBLE PRE-FUSION RSV F POLYPEPTIDES

• US 20160176932A1
• Filed: 06/17/2014
• Published: 06/23/2016
• Est. Priority Date: 06/17/2013
• Status: Active Grant

• Current Assignee: Janssen Vaccines & Prevention B.V. (Johnson & Johnson)
• Original Assignee: Crucell Holland B.V (Johnson & Johnson)
• Inventors: Langedijk, Johannes P.M., Krarup, Anders
• Granted Patent: US 10,294,279 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 31/14 for RNA viruses
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C07K 2319/73 containing coiled-coiled motif (leucine zippers)
• CPC Class Codes: C12N 2760/18522 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
• CPC Class Codes: C12N 2760/18534 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2999/002 Adverse teaching
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

This disclosure provides stable pre-fusion respiratory syncytial virus (RSV) F polypeptides, immunogenic compositions comprising the polypeptides, and uses thereof for the prevention and/or treatment of RSV infection.

Claims

1. A recombinant pre-fusion respiratory syncytial virus (RSV) Fusion (F) polypeptide comprising:

at least one epitope that is specific to the pre-fusion conformation F protein, wherein the at least one epitope is recognized by

a pre-fusion specific monoclonal antibody, antibody comprising;

a heavy chain CDR1 region of SEQ ID NO;

54,

a heavy chain CDR2 region of SEQ ID NO;

55,

a heavy chain CDR3 region of SEQ ID NO;

56,

a light chain CDR1 region of SEQ ID NO;

62,

a light chain CDR2 region of SEQ ID NO;

63, and

a light chain CDR3 region of SEQ ID NO;

64, and/or

a pre-fusion specific monoclonal antibody, antibody comprising;

a heavy chain CDR1 region of SEQ ID NO;

58,

a heavy chain CDR2 region of SEQ ID NO;

59,

a heavy chain CDR3 region of SEQ ID NO;

60,

a light chain CDR1 region of SEQ ID NO;

66,

a light chain CDR2 region of SEQ ID NO;

67, and

a light chain CDR3 region of SEQ ID NO;

68; and

an F1 domain and an F2 domain, said F1 and F2 domains comprising at least one mutation, as compared to wild-type F1 and F2 domains, selected from the group consisting of;

(a) a mutation of the amino acid residue at position 161;

(b) a mutation of the amino acid residue at position 182;

(c) a mutation of the amino acid residue at position 173; and

(d) a mutation of the amino acid residue D at position 486 into C (D486C) in combination with a mutation of the amino acid residue D at position 489 into C (D489C) or a mutation of the amino acid residue E at position 487 into C (E487C).

2. Pre-fusion RSV F polypeptide according to claim 1, wherein the polypeptide is trimeric.

3. The pre-fusion RSV F polypeptide according to claim 1, wherein the least one mutation is selected from the group consisting of:

(a) a mutation of the amino acid residue E at position 161 into P, Q or G (E161P, E161Q) or E161G);

(b) a mutation of the amino acid residue S at position 182 into P (S182P);

(c) a mutation of the amino acid residue S, T or N at position 173 into P (S173P); and

(d) a mutation of the amino acid residue D at position 486 into C (D486C) in combination with a mutation of the amino acid residue D at position 489 into C (D489C) or a mutation of the amino acid residue E at position 487 into C (E487C).

4. The pre-fusion RSV F polypeptide of claim 1, further comprising a mutation of the amino acid residue at position 67 and/or a mutation of the amino acid residue at position 215.

5. The pre-fusion RSV F polypeptide according to claim 4, wherein the polypeptide comprises a mutation of the amino acid residue N or T at position 67 and/or a mutation of amino acid residue S at position 215.

6. The pre-fusion RSV F polypeptide of claim 1, wherein the polypeptide comprises a linking sequence comprising from 1 to 10 amino acids, linking the F1 domain and F2 domain.

7. The pre-fusion RSV F polypeptide of claim 1, wherein the polypeptide comprises a truncated F1 domain.

8. Pre-fusion RSV F polypeptide according to claim 7, wherein the polypeptide comprises a heterologous trimerization domain linked to said truncated F1 domain

9. The pre-fusion RSV F polypeptide according to claim 8, wherein the heterologous trimerization domain comprises GYIPEAPRDGQAYVRKDGEWVLLSTFL (SEQ ID NO:

4).

10. Pre-fusion RSV F polypeptide according to claim 12, wherein the trimerization domain is linked to amino acid residue 513 of the RSV F protein.

11. The pre-fusion RSV F polypeptide of claim 1, wherein the F1 domain and/or the F2 domain are from an RSV A strain.

12. The pre-fusion RSV F polypeptide of claim 1, wherein the F1 domain and/or the F2 domain are from an RSV B strain.

13. The pre-fusion RSV F polypeptide of claim 1, wherein the polypeptide comprises a peptide selected from the group consisting of SEQ ID NO:

90-SEQ ID NO;

94.

14. Nucleic A nucleic acid molecule encoding the pre-fusion RSV F polypeptide of claim 1.

15. The nucleic acid molecule according to claim 14, wherein the nucleic acid molecule has been codon-optimized for expression in mammalian cells.

16. A vector comprising the nucleic acid molecule according to claim 14.

17. A composition comprising:

the pre-fusion RSV F polypeptide of claim 1.

18. A method of inducing an immune response against RSV F protein in a subject, the method comprising:

administering to the subject an amount of the pre-fusion RSV F polypeptide of claim 1 to induce an immune response against RSV F protein in the subject.

19. A vaccine comprising:

the pre-fusion RSV F polypeptide of claim 1.

20. A method of prophylaxing and/or treating an RSV F infection in a subject, the method comprising:

administering to the subject an amount of the pre-fusion RSV F polypeptide of claim 1 to prophylax and/or treat the subject for an RSV infection.

21. A method of inducing an immune response against RSV F protein in a subject, the method comprising:

administering to the subject an amount of the nucleic acid molecule of claim 14 to induce an immune response against RSV F protein in the subject.

22. A vaccine comprising:

the nucleic acid molecule of claim 14.

23. A method of prophylaxing and/or treating an RSV F infection in a subject, the method comprising:

administering to the subject an amount of the nucleic acid molecule of claim 14 to prophylax and/or treat the subject for RSV infection.

24. A host cell transformed with the vector of claim 16.