# US 11,305,003 B2 · Zika virus chimeric polyepitope comprising non-structural proteins and its use in an immunogenic composition

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- Number: US 11,305,003 B2
- Document ID: US11305003B2
- Status: Granted
- Kind: B2
- Application number: 16/762,849
- Filed: 2018-11-08
- Published: 2020-09-10
- Granted: 2022-04-19
- Earliest claimed: 2017-11-09
- Assignees: Institut Pasteur; Universidad El Bosque
- Applicants: Institut Pasteur; Universidad El Bosque
- Inventors: Claude Roth; Etienne Simon-Loriere; Anavaj Sakuntabhai; Felix Delgado
- Web page: https://patentgrep.com/patent/US11305003B2

## CPC

- A61K 39/12: Viral antigens
- C07K 14/1825: {Flaviviruses or Group B arboviruses, e.g. yellow fever virus, japanese encephalitis, tick-borne encephalitis, dengue}
- A61K 9/0029: {Parenteral nutrition; Parenteral nutrition compositions as drug carriers}
- C07K 14/005: from viruses
- 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 2039/70: {Multivalent vaccine}
- C07K 2319/00: Fusion polypeptide
- 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
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- 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)
- 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)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The present invention is directed to a Zika virus (ZIKV) chimeric polyepitope comprising non-structural proteins and its use in an immunogenic composition. The present invention provides means, in particular polynucleotides, vectors and cells expressing said chimeric polyepitope. The present invention also relates to a composition or a vaccine comprising at least one of said polyepitope, polynucleotide, vector or host cell for use in the prevention of a ZIKV infection in a human subject, or for use in the prevention of ZIKV and dengue vims (DENV) infections in a human subject.

## Claims

### Claim 1

A chimeric polyepitope comprising (i) at least the following T-cell epitopes of (a) and (b), or (ii) at least the following T-cell epitopes of (a) and (c), or (iii) at least the following T-cell epitopes of (b) and (c):
(a) a T-cell epitope of the non-structural (NS) NS1 protein of a Zika virus (ZIKV) comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 10-12, 14, 15, 17-19, 23, 24 and 78-83,
(b) a T-cell epitope of the NS3 protein of a ZIKV comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 28, 29, 31, 33-35, 84 and 85,
(c) a T-cell epitope of the NS5 protein of a ZIKV comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 46, 48-50, 52-55, 57-60, 62, 64, 67, 69, 72, 73 and 86-91,
or a T-cell epitope variant thereof, which differs from the original amino acid sequence of the T-cell epitope of (a), (b) or (c) by point mutation of one or more amino acid residues and which has at least 90% sequence identity or more than 95% sequence identity or 99% sequence identity with said original sequence.

### Claim 2 (depends on 1)

The chimeric polyepitope according to claim 1, comprising at least the T-cell epitopes of (a), (b) and (c), or the T-cell epitope variant thereof.

### Claim 3 (depends on 1)

The chimeric polyepitope according to claim 1, consisting of (i) the T-cell epitopes of (a) and (b), or (ii) the T-cell epitopes of (a) and (c), or (iii) the T-cell epitopes of (b) and (c), or (iv) the T-cell epitopes of (a), (b) and (c), or the T-cell epitope variant thereof.

### Claim 4 (depends on 1)

The chimeric polyepitope according to claim 1, wherein the T-cell epitope of (a) comprises or consists of the amino acid sequence selected from the group consisting of SEQ ID NOs: 17, 23 and 78-83, the T-cell epitope of (b) comprises or consists of the amino acid sequence selected from the group consisting of SEQ ID NOs: 31, 33, 84 and 85, and the T-cell epitope of (c) comprises or consists of the amino acid sequence selected from the group consisting of SEQ ID NOs: 46, 48, 52, 57, 62, 64, 67 and 86-91.

### Claim 5 (depends on 1)

The chimeric polyepitope according to claim 1, wherein the T-cell epitope of (a) comprises or consists of the amino acid sequence of SEQ ID NOs: 11, 12, 17-19, 23, 24, 78, 80 and 83, the T-cell epitope of (b) comprises or consists of the amino acid sequence of SEQ ID NOs: 28, 31, 33, 34, 84 and 85, and the T-cell epitope of (c) comprises or consists of the amino acid sequence of SEQ ID NOs: 48-50, 52-55, 57, 58, 60, 62, 67, 88, 89 and 90.

### Claim 6 (depends on 1)

The chimeric polyepitope according to claim 1, further comprising at least one T-cell epitope of a ZIKV protein selected from the group consisting of:
(i) a T-cell epitope of the C protein of a ZIKV comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 4-6 and 75,
(ii) a T-cell epitope of the E protein of a ZIKV comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 7, 76 and 77,
(iii) a T-cell epitope of the NS2B protein of a ZIKV comprising or consisting of the amino acid sequence of SEQ ID NO: 25,
(iv) a T-cell epitope of the NS4A protein of a ZIKV comprising or consisting of the amino acid sequence of SEQ ID NO: 36, and
(v) a T-cell epitope of the NS4B protein of a ZIKV comprising or consisting of the amino acid sequence selected from the group consisting of SEQ ID NOs: 40-43.

### Claim 7 (depends on 6)

The chimeric polyepitope according to claim 6, wherein said at least one T-cell epitope of a ZIKV protein is selected from the group consisting of:
the T-cell epitope of the C protein of a ZIKV comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 4-6 and 75, and
the T-cell epitope of the NS4B protein of a ZIKV comprising or consisting of the amino acid sequence selected from the group consisting of SEQ ID NOs: 40-43.

### Claim 8 (depends on 7)

The chimeric polyepitope according to claim 7, which has an amino acid sequence of SEQ ID NO: 99.

### Claim 9 (depends on 1)

The chimeric polyepitope according to claim 1, which elicits a human leukocyte antigen (HLA)-restricted CD8+ and/or CD4+ T cell response (i) against ZIKV, or (ii) against ZIKV and DENV, in particular DENV serotype 1 (DENV1), DENV serotype 2 (DENV2), DENV serotype 3 (DENV3) and DENV serotype 4 (DENV4).

### Claim 10 (depends on 1)

The chimeric polyepitope according to claim 1, wherein the T-cell epitopes are assembled in a fusion polypeptide.

### Claim 11 (depends on 1)

The chimeric polyepitope according claim 1, wherein the ZIKV is from the African lineage, or from the Asian lineage.

### Claim 12 (depends on 1)

An isolated or purified polynucleotide encoding the chimeric polyepitope according to claim 1.

### Claim 13 (depends on 12)

A vector comprising the polynucleotide according to claim 12.

### Claim 14 (depends on 12)

Transformed eukaryotic cells comprising the polynucleotide according to claim 12.

### Claim 15 (depends on 1)

An immunogenic composition comprising the chimeric polyepitope according to claim 1,
and
an adjuvant and/or a pharmaceutically acceptable vehicle.

### Claim 16 (depends on 15)

The immunogenic composition according to claim 15, wherein said composition is formulated for an administration by a route selected from the group consisting of subcutaneous (s.c.), intradermal (i.d.), intramuscular (i.m.), intraperitoneal (i.p.) and intravenous (i.v.) injection.

### Claim 17 (depends on 15)

The immunogenic composition according to claim 15, formulated for administration in a prime-boost administration regime.

### Claim 18 (depends on 1)

A method of (i) preventing a Zika virus (ZIKV) infection in a human subject or (ii) preventing ZIKV and Dengue virus (DENV) infections in a human subject, comprising administering the chimeric polyepitope according to claim 1 to the human subject.

### Claim 19 (depends on 18)

The method of preventing a ZIKV infection in a human subject according to claim 18, wherein the T-cell epitopes are ZIKV-specific epitopes comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 5, 10, 11, 19, 27, 31, 40-43, 46, 72, 73, 75, 78-80, 82, 84, 85, 87 and 91.

### Claim 20 (depends on 18)

The method of preventing ZIKV and DENV infections in a human subject according to claim 18, wherein the T-cell epitopes are ZIKV-DENV cross-reactive epitopes comprising or consisting of an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 5, 6, 12, 14, 15, 17-19, 23, 24, 27, 28, 33-35, 40, 41, 46, 48-50, 52-55, 57, 59, 60, 62, 64, 67, 69, 72, 73, 84, 85 and 86-91.