# US 7,744,901 B2 · Influenza hemagglutinin and neuraminidase variants

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- Number: US 7,744,901 B2
- Document ID: US7744901B2
- Status: Granted
- Kind: B2
- Application number: 12/354,085
- Filed: 2009-01-15
- Published: 2009-05-28
- Granted: 2010-06-29
- Earliest claimed: 2004-05-25
- Assignees: MedImmune, LLC; National Institute of Health
- Inventors: Chin-Fen Yang; George Kemble; Kanta Subbarao; Brian Murphy
- Web page: https://patentgrep.com/patent/US7744901B2

## CPC

- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- A61K 39/12: Viral antigens
- A61P 31/12: Antivirals
- A61P 31/16: for influenza or rhinoviruses
- A61P 37/04: Immunostimulants
- C07K 14/005: from viruses
- C12Q 1/701: {Specific hybridization probes}
- A61K 2039/5254: {avirulent or attenuated}
- A61K 2039/543: {intranasal}
- A61K 2039/58: {raising an immune response against a target which is not the antigen used for immunisation}
- C12N 2501/70: Enzymes
- C12N 2760/16121: Viruses as such, e.g. new isolates, mutants or their genomic sequences
- C12N 2760/16122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2760/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2760/16161: Methods of inactivation or attenuation
- C12N 2760/16162: by genetic engineering
- C12N 2760/16171: Demonstrated in vivo effect
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES

## Abstract

Polypeptides, polynucleotides, methods, compositions, and vaccines comprising (avian pandemic) influenza hemagglutinin and neuraminidase variants are provided.

## Claims

### Claim 1

A reassortant influenza virus, wherein said virus comprises at least 6 internal genome segments from A/Ann Arbor/6/60 and at least one genome segment comprising a nucleic acid encoding a hemagglutinin polypeptide comprising the amino acid sequence of SEQ ID NO: 11.

### Claim 2 (depends on 1)

An immunogenic composition comprising an immunologically effective amount of the reassortant influenza virus of claim 1.

### Claim 3 (depends on 1)

A method for stimulating the immune system of an individual to produce a protective immune response against influenza virus, the method comprising administering to the individual an immunologically effective amount of the reassortant influenza virus of claim 1 in a physiologically effective carrier.

### Claim 4 (depends on 1)

A method of prophylactic or therapeutic treatment of a viral infection in a subject, the method comprising: administering to the subject, the reassortant virus of claim 1 in an amount effective to produce an immunogenic response against the viral infection.

### Claim 5 (depends on 4)

The method of claim 4, wherein the subject is a human.

### Claim 6 (depends on 2)

The immunogenic composition of claim 2, wherein said reassortant influenza virus further comprises a neuramidinase genome segment comprising a nucleic acid encoding a neuramidinase polypeptide selected from the group consisting of: the polypeptides of SEQ ID NOs: 12, 14, 16, 18, and 20.

### Claim 7 (depends on 2)

A live attenuated influenza vaccine comprising the composition of claim 2.

### Claim 8 (depends on 2)

A split virus or killed virus vaccine comprising the composition of claim 2.

### Claim 9 (depends on 6)

A live attenuated influenza vaccine comprising the composition of claim 6.

### Claim 10 (depends on 6)

A split virus or killed virus vaccine comprising the composition of claim 6.

### Claim 11

A method for producing influenza viruses in cell culture, the method comprising:
i) introducing into a population of host cells a plurality of vectors comprising nucleic acid sequences corresponding to at least 6 internal genome segments of the influenza strain A/Ann Arbor/6/60; and at least one genome segment comprising a nucleic acid encoding a hemagglutinin polypeptide comprising the amino acid sequence of SEQ ID NO: 11,
ii) culturing the population of host cells at a temperature less than or equal to 35° C.; and,
iii) recovering a plurality of influenza viruses.

### Claim 12 (depends on 11)

An immunogenic composition comprising an immunologically effective amount of the influenza virus produced by the method of claim 11.

### Claim 13 (depends on 11)

A method for stimulating the immune system of an individual to produce a protective immune response against influenza virus, the method comprising administering to the individual an immunologically effective amount of the influenza virus produced by the method of claim 11 in a physiologically effective carrier.

### Claim 14 (depends on 12)

A method for stimulating the immune system of an individual to produce a protective immune response against influenza virus, the method comprising administering to the individual the immunogenic composition of claim 12.

### Claim 15 (depends on 12)

A live attenuated influenza vaccine comprising the immunogenic composition of claim 12.

### Claim 16 (depends on 12)

A split virus or killed virus vaccine comprising the immunogenic composition of claim 12.

### Claim 17 (depends on 11)

The method of claim 11, wherein the plurality of vectors further comprise nucleic acids corresponding to a genome segment comprising a nucleic acid encoding a neuramidinase polypeptide comprising the amino acid sequence selected from the group consisting of SEQ ID NOs: 12, 14, 16, 18, and 20.