# US 8,784,839 B2 · Attenuated live vaccines

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- Number: US 8,784,839 B2
- Document ID: US8784839B2
- Status: Granted
- Kind: B2
- Application number: 13/048,412
- Filed: 2011-03-15
- Published: 2011-07-07
- Granted: 2014-07-22
- Earliest claimed: 2001-02-21
- Assignees: Valneva Austria GmbH
- Applicants: Franz Xaver Heinz; Christian Mandl
- Inventors: Franz Xaver Heinz; Christian Mandl
- Web page: https://patentgrep.com/patent/US8784839B2

## CPC

- C07K 14/005: from viruses
- A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
- A61P 31/00: Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P 31/14: for RNA viruses
- A61P 37/04: Immunostimulants
- 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 35/13: Tumour cells, irrespective of tissue of origin (tumour vaccines A61K39/00)
- A61K 2039/5254: {avirulent or attenuated}
- C12N 2770/24122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2770/24134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2770/24161: Methods of inactivation or attenuation
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- 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)

## Abstract

Disclosed is an attenuated flavivirus live vaccine comprising a flavivirus mutant, characterized in that the flavivirus mutant has a deletion in the capsid protein of at least more than 4 successive amino acids, wherein the carboxy-terminal hydrophobic region is not affected by the deletion.

## Claims

### Claim 1

An attenuated tick-borne encephalitis virus (TBE virus) live vaccine comprising a TBE virus mutant, wherein the TBE virus mutant has a deletion in the capsid protein of
(a) 5 to 20 successive amino acids, or
(b) more than 20 successive amino acids, wherein the capsid protein additionally comprises one or more further mutations by which the hydrophobicity of the capsid protein is increased, the further mutation being selected from
i) point mutations by which the hydrophobicity of the capsid protein is increased, and/or
ii) insertions of amino acid sections having predominantly hydrophobic properties, and
wherein the deletion concerns regions of the internal hydrophobic domain of the capsid protein and wherein the carboxy-terminal hydrophobic region is not affected by the deletion.

### Claim 2 (depends on 1)

The TBE virus live vaccine according to claim 1 wherein 5 to 20 or up to 70 successive amino acids in the capsid protein are deleted.

### Claim 3 (depends on 1)

The TBE virus live vaccine according to claim 1, wherein the insertions of amino acid sections having predominantly hydrophobic properties are duplications of hydrophobic regions of the capsid protein.

### Claim 4 (depends on 1)

The TBE virus live vaccine according to claim 1, wherein the vaccine comprises 101 to 107, 102 to 106 or 103 to 105 infectious units of TBE virus.

### Claim 5 (depends on 1)

The TBE virus live vaccine according to claim 1, wherein the vaccine further comprises antibiotics, preservatives, stabilizers, buffer substances or mixtures thereof.

### Claim 6 (depends on 5)

The TBE virus live vaccine according to claim 5, wherein the vaccine comprises neomycin, kanamycin, thimerosal, human albumin, lactose-sorbitol, sorbitol-gelatin, polygeline, MgCl2, MgSO4, amino acids, polysaccharides, buffer salts or mixtures thereof.

### Claim 7 (depends on 1)

The TBE virus live vaccine according to claim 1, wherein the vaccine is produced in non-transformed host cells.

### Claim 8

A tick-borne encephalitis virus (TBE virus) vaccine, wherein the vaccine comprises a nucleic acid which codes for a deleted capsid protein that comprises a deletion of
(a) 5 to 20 successive amino acids, or
(b) more than 20 successive amino acids, wherein the capsid protein additionally comprises one or more further mutations by which the hydrophobicity of the capsid protein is increased, the further mutation being selected from
i) point mutations by which the hydrophobicity of the capsid protein is increased, and/or
ii) insertions of amino acid sections having predominantly hydrophobic properties, and
wherein the deletion concerns regions of the internal hydrophobic domain of the capsid protein and wherein the carboxy-terminal hydrophobic region is not affected by the deletion.

### Claim 9 (depends on 8)

The TBE virus vaccine according to claim 8, wherein the vaccine further comprises aminoglycoside antibiotics, neomycin, kanamycin, liposomes, microspheres or mixtures thereof.

### Claim 10 (depends on 1)

A method of producing an attenuated tick-borne encephalitis virus (TBE virus) live vaccine comprising a TBE virus mutant according to claim 1, comprising the following steps:
providing a TBE virus or a TBE virus nucleic acid, wherein the TBE virus or the TBE virus nucleic acid has a deletion in the capsid protein and further mutations as defined in claim 1,
propagating the TBE virus or the TBE virus nucleic acid in suitable host cells,
recovering the virus particles propagated by the host cells, and
working up the virus particles to a live vaccine.

### Claim 11 (depends on 10)

The method according to claim 10, wherein the host cell is selected from the group consisting of chicken embryo cells, primary chicken embryo cells, human diploid cell lines, vero cells, primary hamster kidney cells, primary canine kidney cells or diploid fetal rhesus lung cells.

### Claim 12 (depends on 2)

The TBE virus live vaccine according to claim 2, wherein 6 to 20 or up to 25 successive amino acids in the capsid protein are deleted.

### Claim 13 (depends on 12)

The TBE virus live vaccine according to claim 12, wherein 7 to 20 successive amino acids in the capsid protein are deleted.