# US 2021/0171979 A1 · Recombinant vectors encoding zika virus protein subunits

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- Number: US 2021/0171979 A1
- Document ID: US20210171979A1
- Status: Published application
- Kind: A1
- Application number: 16/762,836
- Filed: 2018-11-09
- Published: 2021-06-10
- Earliest claimed: 2017-11-10
- Assignees: Research Institute At Nationwide Children's Hospital; Ohio State Innovation Foundation
- Applicants: Research Institute at Nationwide Children's Hospital; Ohio State Innovation Foundation
- Inventors: Mark Peeples; Jianrong Li; Prosper N. Boyaka; Anzhong Li; Mijia Lu; Yuanmei Ma
- Web page: https://patentgrep.com/patent/US20210171979A1

## CPC

- C12N 15/86: Viral vectors
- A61K 39/12: Viral antigens
- A61P 31/14: for RNA viruses
- C07K 14/005: from viruses
- A01K 2227/105: Murine
- A01K 2267/0337: Animal models for infectious diseases
- A61K 2039/5254: {avirulent or attenuated}
- A61K 2039/5256: {expressing foreign proteins}
- A61K 2039/5258: {Virus-like particles}
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K 2039/575: {humoral response}
- C12N 2760/20243: viral genome or elements thereof as genetic vector
- C12N 2760/20262: by genetic engineering
- 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 2770/24171: Demonstrated in vivo effect
- 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
- 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)
- A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Embodiments disclosed herein provide compositions, methods, and uses for recombinant vectors encoding Zika virus (ZIKV) protein subunits, and immunogenic compositions thereof. Certain embodiments provide recombinant vectors encoding ZIKV nonstructural protein 1 (NS 1), and optionally, ZIKV envelope (E) protein and premembrane (prM) protein. Other embodiments provide expression cassettes comprising a promoter operably linked to a polynucleotide that encodes the ZIKV NS 1 protein, and optionally ZIKV E and prM proteins. In some embodiments, the disclosed expression cassettes can be incorporated into a vector to produce a recombinant vector. Also provided are immunogenic compositions comprising one or more recombinant vectors described herein, and methods for inducing an immune response against ZIKV in a subject comprising administering to the subject an immunologically effective dose of an immunogenic composition of the present disclosure.

## Claims

### Claim 1

A recombinant vector comprising a polynucleotide sequence encoding a Zika virus nonstructural protein 1 (NS1 protein).

### Claim 2 (depends on 1)

The recombinant vector of claim 1, wherein the Zika virus NS1 protein has at least 90% amino acid sequence identity with SEQ ID NO: 14.

### Claim 3 (depends on 1)

The recombinant vector of claim 1, wherein the Zika virus NS1 protein comprises an amino acid sequence according to SEQ ID NO: 14.

### Claim 4 (depends on 1)

The recombinant vector of claim 1, wherein the recombinant vector further comprises one or more polynucleotide sequences encoding a Zika virus envelope (E) protein or truncation mutant thereof, and a Zika virus premembrane (prM) protein.

### Claim 5 (depends on 4)

The recombinant vector of claim 4, wherein:
the Zika virus E protein has at least 90% amino acid sequence identity with SEQ ID NO: 4;
the Zika virus E protein truncation mutant has at least 90% amino acid sequence identity with one of SEQ ID NO 10 (E404), SEQ ID NO: 8 (E414), or SEQ ID NO: 6 (E415); and
the Zika virus prM protein has at least 90% amino acid sequence identity with SEQ ID NO: 12.

### Claim 6 (depends on 4)

The recombinant vector of claim 4, wherein the recombinant vector encodes the Zika virus NS1 protein, the Zika virus E protein or truncation mutant thereof, and the Zika virus prM protein.

### Claim 7 (depends on 1)

The recombinant vector of claim 1, wherein the recombinant vector comprises a DNA plasmid vector or an RNA viral vector.

### Claim 8 (depends on 7)

The recombinant vector of claim 7, wherein the viral vector is selected from the group comprising adenovirus, adeno-associated virus (AAV), retrovirus, lentivirus, vaccinia virus, cytomegalovirus, Sendai virus, modified vaccinia Ankara virus, and vesicular stomatitis virus (VSV).

### Claim 9 (depends on 1)

The recombinant vector of claim 1, wherein the recombinant vector comprises a VSV vector.

### Claim 10 (depends on 9)

The recombinant vector of claim 9, wherein the VSV vector comprises at least one mutation in a methyltransferase-encoding region of an L protein of the VSV vector.

### Claim 11 (depends on 10)

The recombinant vector of claim 10, wherein the at least one mutation is a nucleic acid mutation that results in an amino acid mutation at a position in the VSV vector selected from the group of K1651, G1670, D1762, K1795, and E1833.

### Claim 12 (depends on 10)

The recombinant vector of claim 10, wherein the at least one mutation is a nucleic acid mutation that results in a G1670A mutation or a D1762A mutation in the VSV vector.

### Claim 13 (depends on 9)

The recombinant vector of claim 9, wherein the VSV vector comprises a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 16.

### Claim 14 (depends on 9)

The recombinant vector of claim 9, wherein the VSV vector comprises a nucleic acid sequence according to SEQ ID NO: 16, or SEQ ID NO: 16 encoding a G→A mutation at amino acid position 1670 of VSV L protein, or SEQ ID NO: 16 encoding a D→A mutation at amino acid position 1762 of VSV L protein.

### Claim 15 (depends on 1)

An immunogenic composition comprising at least one recombinant vector according to claim 1 and a pharmaceutically acceptable excipient.

### Claim 16 (depends on 15)

The immunogenic composition of claim 15, further comprising an adjuvant.

### Claim 17 (depends on 15)

A method for inducing an effective immune response against Zika virus in a subject, the method comprising administering to the subject an immunologically effective dose of the immunogenic composition of claim 15.

### Claim 18 (depends on 17)

The method of claim 17, wherein the subject is human.

### Claim 19 (depends on 18)

The method of claim 18, wherein the subject is pregnant, may be pregnant, or is trying to get pregnant.

### Claim 20 (depends on 17)

The method of claim 17, wherein the immunogenic composition is administered to the subject via a route selected from intranasal administration, subcutaneous administration, intramuscular administration, intradermal administration, and oral administration.

### Claim 21 (depends on 17)

The method of claim 17, further comprising administering at least one subsequent immunologically effective dose of the immunogenic composition.

### Claim 22

A method for inducing an effective immune response against Zika virus in a subject, the method comprising expressing a Zika virus nonstructural protein 1 (NS1 protein) in cells of the subject.

### Claim 23 (depends on 22)

The method of claim 22, wherein the Zika virus NS1 protein has at least 90% amino acid sequence identity with SEQ ID NO: 14.

### Claim 24 (depends on 22)

The method of claim 22, wherein the Zika virus NS1 protein comprises an amino acid sequence according to SEQ ID NO: 14.

### Claim 25 (depends on 22)

The method of claim 22, further comprising co-expressing a Zika virus envelope (E) protein or a truncation mutant thereof, and a Zika virus premembrane (prM) protein.

### Claim 26 (depends on 25)

The method of claim 25, wherein:
the Zika virus E protein has at least 90% amino acid sequence identity with SEQ ID NO: 4;
the Zika virus E protein truncation mutant has at least 90% sequence identity with one of SEQ ID NO 10 (E404), SEQ ID NO: 8 (E414), or SEQ ID NO: 6 (E415); and
the Zika virus prM protein has at least 90% amino acid sequence identity with SEQ ID NO: 12.

### Claim 27 (depends on 22)

The method of claim 22, wherein the Zika virus protein(s) are expressed from a recombinant vesicular stomatitis virus (VSV) vector.

### Claim 28

An expression cassette comprising a promoter operably linked to a polynucleotide encoding a Zika virus nonstructural protein 1 (NS1 protein).

### Claim 29 (depends on 28)

The expression cassette of claim 28, wherein the polynucleotide encoding the Zika virus NS1 protein further encodes a Zika virus envelope (E) protein or a truncation mutant thereof, and a Zika virus premembrane (prM) protein.

### Claim 30 (depends on 29)

The expression cassette of claim 29, wherein the Zika virus NS1 protein has at least 90% amino acid sequence identity with SEQ ID NO: 14, the Zika virus E protein has at least 90% amino acid sequence identity with SEQ ID NO: 4, the Zika virus E protein truncation mutant has at least 90% sequence identity with one of SEQ ID NO 10 (E404), SEQ ID NO: 8 (E414), or SEQ ID NO: 6 (E415), and the Zika virus prM protein has at least 90% amino acid sequence identity with SEQ ID NO: 12.