# US 10,369,211 B2 · Influenza H5 vaccines

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,369,211 B2
- Document ID: US10369211B2
- Status: Granted
- Kind: B2
- Application number: 15/662,358
- Filed: 2017-07-28
- Published: 2017-12-07
- Granted: 2019-08-06
- Earliest claimed: 2011-08-15
- Applicants: Boehringer Ingelheim Vetmedica GmbH; Boehringer Ingelheim Vetmedica S.A. de C.V.
- Inventors: Mauricio Realpe-Quintero; Paulino Carlos Gonzalez-Hernandez; Eric Vaughn
- Web page: https://patentgrep.com/patent/US10369211B2

## CPC

- A61K 39/145: Orthomyxoviridae, e.g. influenza virus
- A61K 39/12: Viral antigens
- A61K 39/17: Newcastle disease virus
- A61P 31/12: Antivirals
- A61P 31/16: for influenza or rhinoviruses
- C07K 14/005: from viruses
- A61K 2039/5256: {expressing foreign proteins}
- A61K 2039/55555: {Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers}
- A61K 2039/55566: {Emulsions, e.g. Freund's adjuvant, MF59}
- A61K 2039/70: {Multivalent vaccine}
- C12N 2710/14043: viral genome or elements thereof as genetic vectore
- C12N 2760/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2760/18134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention is based on the surprising finding that H5 protein of clade 1 H5N1 induces, in particular by a single-shot vaccination, a cross-clade protective immune response to influenza viruses with H5N1 HA. In one aspect, the invention is thus directed to H5 protein of clade 1 H5N1 virus for use in a method of treating or preventing infections with H5N1 virus of a different clade, namely of a clade different from clade 1 or from any clade with the exception of clade 1, respectively.

## Claims

### Claim 1

A method for protecting an animal against a clade 2 H5N1 influenza virus infection, comprising:
administering to the animal a composition comprising (i) a crude whole cell H5 protein of a clade 1 H5N1 influenza virus, expressed in insect cells by a baculovirus based expression system, in an amount effective to induce a cross-clade protective immune response in the animal against the clade 2 H5N1 influenza virus infection, and (ii) an inactivated Newcastle disease virus;
wherein the H5 protein of the clade 1 H5N1 influenza virus comprises a polypeptide sequence that has at least 98% sequence identity to SEQ ID NO:1;
wherein the H5 protein of the clade 1 H5N1 influenza virus has amino acid 223N and modification 328K+;
wherein amino acid position numbering of the H5 protein of the clade 1 H5N1 influenza virus refers to amino acid positions as given in SEQ ID NO:2; and
wherein modification 328K+ means that a second Lysine (K+) is inserted at amino acid position 328 of the H5 protein of the clade 1 H5N1 influenza virus.

### Claim 2 (depends on 1)

The method of claim 1, wherein the H5 protein of the clade 1 H5N1 influenza virus has amino acid 94N, amino acid 120N, amino acid 155N, or any combinations thereof.

### Claim 3 (depends on 1)

The method of claim 1, wherein the H5 protein of the clade 1 H5N1 influenza virus comprises an amino acid sequence according to:
i. any one of SEQ ID NOs: 5-7;
ii. any one of SEQ ID NOs: 5-7 modified to have at least one of the following amino acid clusters:
a. aa 93-95: GNF
b. aa 123-125: SDH
c. aa 128-130: SSG
d. aa 138-140: GSS
e. aa 226-228: MDF
f. aa 270-272: EVE
g. aa 309-311: NKL;
iii. any one of SEQ ID NOs: 5-7 modified to have at least one of the following amino acids: 36T, 36K, 83A, 83T, 83D, 86A, 86V, 120S, 155S, 156A, 156T, 189R, 189K, 212K, 212R, 212E, 263A, or 263T; or
iv. any one of SEQ ID NOs: 5-7 modified to have at least one of the following amino acids: 36T, 36K, 83A, 83T, 83D, 86A, 86V, 120S, 155S, 156A, 156T, 189R, 189K, 212K, 212R, 212E, 263A, or 263T, and modified to have at least one of the following amino acid clusters:
a. aa 93-95: GNF
b. aa 123-125: SDH
c. aa 128-130: SSG
d. aa 138-140: GSS
e. aa 226-228: MDF
f. aa 270-272: EVE
g. aa 309-311: NKL.

### Claim 4 (depends on 1)

The method of claim 1, wherein the clade 2 H5N1 influenza virus is a clade 2.2 or clade 2.3 H5N1 influenza virus.

### Claim 5 (depends on 1)

The method of claim 1, wherein the clade 2 H5N1 influenza virus is a clade 2.2.1 or clade 2.3.2 H5N1 influenza virus.

### Claim 6 (depends on 1)

The method of claim 1, wherein the clade 2 H5N1 influenza virus is of North African or Vietnamese origin.

### Claim 7 (depends on 1)

The method of claim 1, wherein the clade 2 H5N1 influenza virus comprises an H5 protein having:
i. amino acids 113D, 126H, 145(−), 156R, 160F, 167T, and 181N, wherein modification 145(−) means that amino acid position 145 is deleted;
ii. amino acids 87P, 145L, 172T, 201E, 206I, 208K, 254T, 341G, and 421K; or
iii. amino acids 145L, 172T, and 254V,
wherein amino acid position numbering of the H5 protein of the clade 2 H5N1 influenza virus refers to amino acid positions as given in SEQ ID NO:8.

### Claim 8 (depends on 1)

The method of claim 1, wherein the clade 2 H5N1 influenza virus comprises an H5 protein having at least 95% sequence identity with at least one of SEQ ID NOs: 9-46.

### Claim 9 (depends on 1)

The method of claim 1, wherein the clade 2 H5N1 influenza virus comprises an H5 protein having:
i. amino acids 87L, 113D, 126H, 145(−), 156R, 160F, 167T, and 181N;
ii. amino acids 87P, 113N, 126R, 145L, 160Y, 172T, 181H, 201E, 206I, 208K, 254T, 341G and 421K, or
iii. amino acids 87L, 113N, 126R, 145L, 156G, 160Y, 172T, 181H, and 254V,
wherein amino acid position numbering of the H5 protein of the clade 2 H5N1 influenza virus refers to amino acid positions as given in SEQ ID NO:8.

### Claim 10 (depends on 1)

The method of claim 1, wherein the inactivated Newcastle disease virus was obtained by inactivating a Newcastle disease virus comprising a RNA polynucleotide having at least 95% sequence identity with a RNA copy of SEQ ID NO: 51.

### Claim 11 (depends on 1)

The method of claim 1, wherein the inactivated Newcastle disease virus is an inactivated LaSota strain Newcastle disease virus.

### Claim 12 (depends on 1)

The method of claim 1, wherein the inactivated Newcastle disease virus was obtained by inactivation with formaldehyde, binary ethylenimine, β-propiolactone, or any combinations thereof.

### Claim 13 (depends on 1)

The method of claim 1, wherein the animal is a poultry.

### Claim 14 (depends on 1)

The method of claim 1, wherein the composition is administered as a single dose.

### Claim 15

A method for protecting an animal against clade 2 H5N1 influenza virus infection, comprising:
administering to the animal a vaccine comprising (i) an H5 protein of a clade 1 H5N1 influenza virus in an amount effective to induce a cross-clade protective immune response in the animal against clade 2 H5N1 influenza virus infection, (ii) an inactivated Newcastle disease virus, and (iii) an adjuvant,
wherein the H5 protein of the clade 1 H5N1 influenza virus comprises a polypeptide sequence that has at least 98% sequence identity to SEQ ID NO:1,
wherein amino acid position numbering of the H5 protein of the clade 1 H5N1 influenza virus refers to amino acid positions as given in SEQ ID NO:2; and
wherein modification 328K+ means that a second Lysine (K+) is inserted at amino acid position 328 of the H5 protein of the clade 1 H5N1 influenza virus.

### Claim 16 (depends on 15)

The method of claim 15, wherein the adjuvant is an oil-in water emulsified adjuvant.

### Claim 17 (depends on 15)

The method of claim 15, wherein the H5 protein of the clade 1 H5N1 influenza virus has amino acid 94N, amino acid 120N, amino acid 155N, or any combinations thereof.

### Claim 18 (depends on 15)

The method of claim 15, wherein the H5 protein of the clade 1 H5N1 influenza virus comprises an amino acid sequence according to:
i. any one of SEQ ID NOs: 5-7;
ii. any one of SEQ ID NOs: 5-7 modified to have at least one of the following amino acid clusters:
a. aa 93-95: GNF
b. aa 123-125: SDH
c. aa 128-130: SSG
d. aa 138-140: GSS
e. aa 226-228: MDF
f. aa 270-272: EVE
g. aa 309-311: NKL;
iii. any one of SEQ ID NOs: 5-7 modified to have at least one of the following amino acids: 36T, 36K, 83A, 83T, 83D, 86A, 86V, 120S, 155S, 156A, 156T, 189R, 189K, 212K, 212R, 212E, 263A, or 263T; or
iv. any one of SEQ ID NOs: 5-7 modified to have at least one of the following amino acids: 36T, 36K, 83A, 83T, 83D, 86A, 86V, 120S, 155S, 156A, 156T, 189R, 189K, 212K, 212R, 212E, 263A, or 263T, and modified to have at least one of the following amino acid clusters:
a. aa 93-95: GNF
b. aa 123-125: SDH
c. aa 128-130: SSG
d. aa 138-140: GSS
e. aa 226-228: MDF
f. aa 270-272: EVE
g. aa 309-311: NKL.

### Claim 19 (depends on 15)

The method of claim 15, wherein the clade 2 H5N1 influenza virus is a clade 2.2 or clade 2.3 H5N1 influenza virus.

### Claim 20 (depends on 15)

The method of claim 15, wherein the clade 2 H5N1 influenza virus is a clade 2.2.1 or clade 2.3.2 H5N1 influenza virus.

### Claim 21 (depends on 15)

The method of claim 15, wherein the clade 2 H5N1 influenza virus is of North African or Vietnamese origin.

### Claim 22 (depends on 15)

The method of claim 15, wherein the clade 2 H5N1 influenza virus comprises an H5 protein having:
i. amino acids 113D, 126H, 145(−), 156R, 160F, 167T, and 181N, wherein modification 145(−) means that amino acid position 145 is deleted;
ii. amino acids 87P, 145L, 172T, 201E, 206I, 208K, 254T, 341G, and 421K; or
iii. amino acids 145L, 172T, and 254V,
wherein amino acid position numbering of the H5 protein of the clade 2 H5N1 influenza virus refers to amino acid positions as given in SEQ ID NO:8.

### Claim 23 (depends on 15)

The method of claim 15, wherein the clade 2 H5N1 influenza virus comprises an H5 protein having at least 95% sequence identity with at least one of SEQ ID NOs: 9-46.

### Claim 24 (depends on 15)

The method of claim 15, wherein the clade 2 H5N1 influenza virus comprises an H5 protein having:
i. amino acids 87L, 113D, 126H, 145(−), 156R, 160F, 167T, and 181N;
ii. amino acids 87P, 113N, 126R, 145L, 160Y, 172T, 181H, 201E, 206I, 208K, 254T, 341G and 421K, or
iii. amino acids 87L, 113N, 126R, 145L, 156G, 160Y, 172T, 181H, and 254V,
wherein amino acid position numbering of the H5 protein of the clade 2 H5N1 influenza virus refers to amino acid positions as given in SEQ ID NO:8.

### Claim 25 (depends on 15)

The method of claim 15, wherein the inactivated Newcastle disease virus was obtained by inactivating a Newcastle disease virus comprising a RNA polynucleotide having at least 95% sequence identity with a RNA copy of SEQ ID NO: 51.

### Claim 26 (depends on 15)

The method of claim 15, wherein the inactivated Newcastle disease virus is an inactivated LaSota strain Newcastle disease virus.

### Claim 27 (depends on 15)

The method of claim 15, wherein the inactivated Newcastle disease virus was obtained by inactivation with formaldehyde, binary ethylenimine, β-propiolactone, or any combinations thereof.

### Claim 28 (depends on 15)

The method of claim 15, wherein the animal is a poultry.

### Claim 29 (depends on 15)

The method of claim 15, wherein the composition is administered as a single dose.