# US 10,117,925 B2 · Influenza virus vaccines and uses thereof

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- Number: US 10,117,925 B2
- Document ID: US10117925B2
- Status: Granted
- Kind: B2
- Application number: 15/324,939
- Filed: 2015-07-09
- Published: 2017-07-06
- Granted: 2018-11-06
- Earliest claimed: 2014-07-10
- Assignees: Janssen Vaccines & Prevention B.V.
- Applicants: Janssen Vaccines & Prevention B.V.
- Inventors: Antonietta Impagliazzo; Jan Willem Meijberg; Katarina Radosevic; Michelle Wagner; Zhaoqing Ding
- Web page: https://patentgrep.com/patent/US10117925B2

## CPC

- A61K 39/145: Orthomyxoviridae, e.g. influenza virus
- A61K 39/12: Viral antigens
- A61P 31/16: for influenza or rhinoviruses
- A61P 37/04: Immunostimulants
- C07K 14/11: Orthomyxoviridae, e.g. influenza virus
- A61K 2039/55577: {Saponins; Quil A; QS21; ISCOMS}
- C12N 2760/16122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2760/16123: Virus like particles [VLP]
- C12N 2760/16134: 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

Provided herein are multimeric influenza hemagglutinin stem domain polypeptides, methods for providing hemagglutinin stem domain polypeptides, compositions comprising the same, vaccines comprising the same and methods of their use, in particular in the detection, prevention and/or treatment of influenza.

## Claims

### Claim 1

A multimeric influenza hemagglutinin stem domain polypeptide, wherein the multimeric influenza hemagglutinin stem domain polypeptide comprises at least a first and a second influenza hemagglutinin stem domain monomer, said first and second monomer each comprising: (a) an influenza hemagglutinin HA1 domain that comprises an HA1 amino (N)-terminal stem segment, wherein said HA1 N-terminal segment comprises the amino acids 1-52 of HA1, preferably the amino acids 18-52 of HA1, covalently linked by a linking sequence of less than 10 residues to an HA1 carboxy (C)-terminal stem segment, comprising the amino acids from position 321 to the C-terminal amino acid of HA1, wherein said HA1 C-terminal segment is linked to (b) an influenza hemagglutinin HA2 domain, and
(c) wherein the polypeptide comprises no protease cleavage site at the junction between the HA1 and HA2 domains; and
(d) wherein the first monomer is linked to said second monomer by a disulfide bridge between the amino acid on position 411 of the first monomer and the amino acid on position 419 of the second monomer, and wherein in each influenza hemagglutinin stem domain monomer the region comprising the amino acid residues 402-418 comprises the amino acid sequence X1NTQX2TAX3GKEX4N(H/K)X5E(K/R) (SEQ ID NO: 8),
wherein X1 is M, X2 is Y, X3 is I, X4 is Y and X5 is S; and
wherein in each influenza hemagglutinin stem domain monomer the amino acid sequence CMKQIEDKIEEIESK (SEQ ID NO: 193) has been introduced at positions 419-433; and
wherein the amino acid residue on position 337 is K; the amino acid residue on position 340 is K, the amino acid residue on position 352 is F and the amino acid residue on position 353 is T;
wherein the HA1 and HA2 domains are derived from an influenza A virus subtype derived from phylogenetic group 1, preferably from an influenza A virus subtype comprising HA of the H1 subtype, and wherein the numbering of the amino acid positions is in relation to SEQ ID NO: 1.

### Claim 2 (depends on 1)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 1, wherein the multimeric polypeptide is trimeric.

### Claim 3 (depends on 1)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 1, wherein the HA2 domain of each influenza hemagglutinin stem domain monomer has been truncated.

### Claim 4 (depends on 3)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 3, wherein the C-terminal part of the HA2 domain from position 519 to the C-terminal amino acid has been deleted.

### Claim 5 (depends on 3)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 3, wherein the C-terminal part of the HA2 domain from position 530 to the C-terminal amino acid has been deleted.

### Claim 6 (depends on 3)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 3, wherein the C-terminal part of the HA2 domain has been replaced by the amino acid sequence GYIPEAPRDGQAYVRKDGEWVLLSTFL (SEQ ID NO: 3), optionally connected through a linker.

### Claim 7 (depends on 1)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 1, wherein the C- terminal amino acid residue of the HA1 C-terminal stem segment of each influenza hemagglutinin stem domain monomer is any amino acid other than arginine (R) or lysine (K), preferably glutamine (Q).

### Claim 8 (depends on 1)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 1, wherein the polypeptide comprises one or more further mutations in the HA1domain and/or the HA2 domains.

### Claim 9 (depends on 1)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 1, wherein the polypeptide selectively binds to the antibodies CR6261 and/or CR9114.

### Claim 10 (depends on 1)

A nucleic acid molecule encoding the polypeptide of claim 1.

### Claim 11 (depends on 10)

A vector comprising the nucleic acid molecule of claim 10.

### Claim 12 (depends on 1)

A composition comprising the polypeptide according to claim 1 and a pharmaceutically acceptable carrier.

### Claim 13 (depends on 10)

A composition comprising the nucleic acid molecule according to claim 10 and a pharmaceutically acceptable carrier.

### Claim 14 (depends on 1)

A method of inducing an immune response against influenza virus in a subject in need thereof, the method comprising administering to the subject in need thereof the polypeptide according to claim 1.

### Claim 15 (depends on 10)

A method of inducing an immune response against influenza in a subject in need thereof, the method comprising administering to the subject in need thereof the nucleic acid molecule according to claim 10.

### Claim 16 (depends on 11)

A method of inducing an immune response against influenza in a subject in need hereof, the method comprising administering to the subject in need thereof the vector according to claim 11.

### Claim 17 (depends on 11)

A composition comprising the vector according to claim 11 and a pharmaceutically acceptable carrier.

### Claim 18 (depends on 1)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 1, wherein each polypeptide monomer comprises the amino acid sequence:
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMVTGLRNX1PSX2QSQGLFGAIAGX3X4EGGWTGMVDGWYGYHHQNEQGSGYAADQKSTQNAINGITNKVNSVIEKX5NTQX6TAX7GCEX8NKX9ERCMKQIEDKIEEIESKIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNNAKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDG (SEQ ID NO: 146), wherein X1 is K, X2 is K, X3 is F, X4 is T, X5 is M, X6 is Y, X7 is I, X8 is Y and X9 is S.

### Claim 19 (depends on 1)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 1, wherein each polypeptide monomer comprises the amino acid sequence:
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMVTGLRNX1PSX2QSQGLFGAIAGX3X4EGGWTGMVDGWYGYHHQNEQGSGYAADQKSTQNAINGITNKVNSVIEKX5NTQX6TAX7GCEX8NKX9ERCMKQIEDKIEEIESKIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNNAKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDGVKLESMGVYQIEG (SEQ ID NO: 147), wherein X1 is K, X2 is K, X3 is F, X4 is T, X5 is M, X6 is Y, X7 is I, X8 is Y and X9 is S.

### Claim 20 (depends on 1)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 1, wherein each polypeptide monomer comprises the amino acid sequence:
DTICIGYHANNSTDTVDTVLEKNVTVTHSVNLLENGGGGKYVCSAKLRMVTGLRNX1PSX2QSQGLFGAIAGX3X4EGGWTGMVDGWYGYHHQNEQGSGYAADQKSTQNAINGITNKVNSVIEKX5NTQX6TAX7GCEX8NKX9ERCMKQIEDKIEEIESKIWCYNAELLVLLENERTLDFHDSNVKNLYEKVKSQLKNNAKEIGNGCFEFYHKCNDECMESVKNGTYDYPKYSEESKLNREKIDGVKLESMGVYQILAIYSTVASSLVLLVSLGAISFWMCSNGSLQCRICI (SEQ ID NO: 148), wherein X1 is K, X2 is K, X3 is F, X4 is T, X5 is M, X6 is Y, X7 is I, X8 is Y and X9 is S.

### Claim 21 (depends on 1)

The multimeric influenza hemagglutinin stem domain polypeptide according to claim 1, wherein each polypeptide monomer comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 164, 186, 202, 203, 207-216, 218-226, and 254-258.