AAV mediated influenza vaccines

Patent metadata

Field	Value
Publication number	US-10786568-B2
Application number	US-201815906887-A
Country	US
Kind code	B2
Filing date	Feb 27, 2018
Priority date	Feb 28, 2017
Publication date	Sep 29, 2020
Grant date	Sep 29, 2020

Abstract

A non-replicating recombinant adeno-associated associated virus (rAAV) having an AAV capsid having packaged therein a vector genome which comprises AAV inverted terminal repeat sequences and at least one nucleic acid sequence encoding four different immunoglobulin regions (a), (b), (c) and (d) is provided. The rAAV-expressed immunoglobulins are useful for providing passive immunization against influenza A and influenza B. Also described herein are compositions containing the rAAV. Methods of vaccinating patients against influenza are provided.

First claim

The invention claimed is:

1. A non-replicating recombinant adeno-associated virus (rAAV) comprising an AAV capsid and a vector genome packaged in the AAV capsid, wherein the vector genome comprises AAV inverted terminal repeat sequences and an expression cassette comprising a nucleic acid sequence encoding an anti-influenza fusion protein having the amino acid sequence of SEQ ID NO: 27.

2. The rAAV according to claim 1 , wherein the vector genome further comprises a 5′ UTR which comprises is-a fragment of human c-myc 5′ UTR.

3. The rAAV according to claim 1 , wherein the vector genome comprises a single-stranded AAV 5′ inverted terminal repeat and an AAV 3′ inverted terminal repeat.

4. The rAAV according to claim 1 , wherein the nucleic acid sequence encoding the anti-influenza fusion protein is of: SEQ ID NO: 21, or SEQ ID NO: 26.

5. The rAAV according to claim 1 , wherein the rAAV has an AAVhu68 capsid, wherein the AAVhu68 capsid comprises:
(a) a heterogenous population of AAVhu68 vp1 proteins selected from one or more of: vp1 proteins produced by expression from a nucleic acid sequence which encodes the predicted amino acid sequence of 1 to 736 of SEQ ID NO:16, vp1 proteins produced from SEQ ID NO: 18, or vp1 proteins produced from a nucleic acid sequence at least 70% identical to SEQ ID NO:18 which encodes the predicted amino acid sequence of 1 to 736 of SEQ ID NO:16;
(b) a heterogenous population of AAVhu68 vp2 proteins selected from one or more of: vp2 proteins produced by expression from a nucleic acid sequence which encodes the predicted amino acid sequence of at least about amino acids 138 to 736 of SEQ ID NO:16, vp2 proteins produced from a sequence comprising at least nucleotides 412 to 2211 of SEQ ID NO:18, or vp2 proteins produced from a nucleic acid sequence at least 70% identical to at least nucleotides 412 to 2211 of SEQ ID NO:18 which encodes the predicted amino acid sequence of at least about amino acids 138 to 736 of SEQ ID NO:16; and
(c) a heterogenous population of AAVhu68 vp3 proteins selected from one or more of: vp3 proteins produced by expression from a nucleic acid sequence which encodes the predicted amino acid sequence of at least about amino acids 203 to 736 of SEQ ID NO:16, vp3 proteins produced from a sequence comprising at least nucleotides 607 to 2211 of SEQ ID NO:18, or vp3 proteins produced from a nucleic acid sequence at least 70% identical to at least nucleotides 607 to 2211 of SEQ ID NO:18 which encodes the predicted amino acid sequence of at least about amino acids 203 to 736 of SEQ ID NO:16.

6. The rAAV according to claim 5 , wherein the nucleic acid sequence encoding the vp1 proteins, vp2 proteins, and vp3 proteins is SEQ ID NO: 18, or a sequence at least 80% to at least 99% identical to SEQ ID NO: 18 which encodes the amino acid sequence of SEQ ID NO:16.

7. The rAAV according to claim 6 , wherein the sequence is at least 80% to 97% identical to SEQ ID NO: 18.

8. The rAAV according to claim 1 , wherein the rAAV has an AAV9 capsid, wherein said AAV9 capsid comprises vp1 capsid proteins having an amino acid sequence of SEQ ID NO: 17 or an amino acid sequence encoded by SEQ ID NO: 36.

9. A composition comprising a carrier, diluent or excipient and a stock of at least the non-replicating rAAV according to claim 1 .

10. The composition according to claim 9 , wherein the composition is formulated for intranasal administration.

11. The composition according to claim 9 , wherein the composition is formulated for intramuscular or intravenous administration.

12. The A method for immunizing a human against influenza, wherein the method comprises intranasally administering to a human in need thereof about 10 9 genome copies (GC) to about 10 14 GC of the rAAV according to claim 1 .

13. A product which comprises a container comprising the rAAV according to claim 1 , optional diluent, and instructions for administration.

14. The rAAV according to claim 1 , wherein the expression cassette comprises a nucleic acid sequence of SEQ ID NO: 26.

15. The rAAV according to claim 1 , wherein the expression cassette further comprises a chicken beta actin promoter.

16. The rAAV according to claim 1 , wherein the expression cassette further comprises a rabbit beta globin polyA.

17. The rAAV according to claim 1 , wherein the expression cassette comprises the sequence of nucleotides 1992 to 4298 of SEQ ID NO: 5.

18. The composition according to claim 9 , wherein the composition is formulated for delivery to the lung.

Assignees

• Univ Pennsylvania
• Janssen Biotech Inc

Inventors

• Limberis Maria P
• Tretiakova Anna P
• Wilson James M
• Naso Michael
• Kolkman Joost
• Friesen Robert
• Wang Qiang

Classifications

• C07K16/108

  Orthomyxoviridae (F), e.g. influenza virus · CPC title

• C12N2840/203

  having an IRES · CPC title

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• C12N2750/14121

  Viruses as such, e.g. new isolates, mutants or their genomic sequences · CPC title

• C12N7/00 Primary

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