# US 2019/0314482 A1 · Vaccines against infectious diseases caused by positive stranded rna viruses

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- Number: US 2019/0314482 A1
- Document ID: US20190314482A1
- Status: Published application
- Kind: A1
- Application number: 16/462,383
- Filed: 2017-11-21
- Published: 2019-10-17
- Earliest claimed: 2016-11-28
- Assignees: Medigen, Inc.
- Applicants: Medigen, Inc.
- Inventors: Peter Pushko; Irina Tretyakova
- Web page: https://patentgrep.com/patent/US20190314482A1

## CPC

- A61K 39/12: Viral antigens
- A61P 31/14: for RNA 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)
- C12N 15/86: Viral vectors
- A61K 2039/5254: {avirulent or attenuated}
- A61K 2039/53: {DNA (RNA) vaccination}
- A61K 2039/552: {Veterinary vaccine}
- A61K 2039/70: {Multivalent vaccine}
- C12N 2310/12: catalytic nucleic acids, e.g. ribozymes
- C12N 2770/24121: Viruses as such, e.g. new isolates, mutants or their genomic sequences
- C12N 2770/24134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2770/24143: viral genome or elements thereof as genetic vector
- C12N 2770/24144: Chimeric viral vector comprising heterologous viral elements for production of another viral vector
- C12N 2770/24151: Methods of production or purification of viral material
- C12N 2830/42: being an intron or intervening sequence for splicing and/or stability of RNA
- 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)
- 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)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Compositions for protecting subjects from diseases caused by (+) SS RNA virus are described herein. The compositions include (i) a vector containing a DNA encoding a RNA molecule of an infectious (+) SS RNA virus operably linked to a eukaryotic RNA polymerase promoter and a carrier; or (ii) (+) SS RNA viruses obtained from eukaryotic cells transfected with the vector of (i) and a carrier.

## Claims

### Claim 1

A vector comprising a DNA encoding a RNA molecule operably linked to a promoter suitable for expression of the DNA in a eukaryotic cell, wherein the RNA molecule encodes an infectious positive single stranded ((+)SS) RNA virus, and wherein the DNA comprises at least three introns.

### Claim 2 (depends on 1)

The vector of claim 1, wherein the (+)SS RNA virus is a flavivirus, alphavirus, picornavirus, rubivirus, coronavirus, Norwalk virus, Hepatitis A virus, Hepatitis C virus, severe acute respiratory (SAR) virus, and lentivirus.

### Claim 3 (depends on 2)

The vector of claim 2, wherein the flavivirus is a Japanese encephalitis virus (JEV), Dengue virus, Yellow Fever virus, West Nile virus, tick borne encephalitis virus, Hepatitis C virus, and Zika virus.

### Claim 4 (depends on 3)

The vector of claim 3, wherein the flavivirus is JEV.

### Claim 5

A vector comprising a DNA encoding a chimeric RNA molecule operably linked to a promoter suitable for expression of the DNA in a eukaryotic cell, wherein the chimeric RNA molecule encodes an infectious (+)SS RNA virus, and wherein the DNA comprises at least three introns.

### Claim 6 (depends on 5)

The vector of claim 5, wherein the (+)SS RNA virus is encoded by a chimeric RNA molecule comprising the RNA of at least two different (+)SS RNA viruses.

### Claim 7 (depends on 6)

The vector of claim 6, wherein the at least two different (+)SS RNA viruses comprise JEV and at least one other virus, Yellow fever virus and at least one other virus, or Dengue virus and at least one other virus.

### Claim 8 (depends on 7)

The vector of claim 7, wherein the at least one other virus is a Zika virus, a West Nile virus, a Yellow fever virus, or a Dengue virus.

### Claim 9 (depends on 8)

The vector of claim 8, wherein the at least two different (+)SS RNA viruses are JEV and Zika virus.

### Claim 10 (depends on 1)

The vector of claim 1, wherein at least one intron is in a region encoding a non-structural protein and one intron is in a region encoding a structural protein.

### Claim 11 (depends on 10)

The vector of claim 10, wherein the intron contains a stop codon or several stop codons.

### Claim 12 (depends on 11)

The vector of claim 11, wherein the DNA comprises three introns.

### Claim 13 (depends on 1)

The vector of claim 1, wherein the DNA comprises at least four introns, at least five introns, or at least six introns.

### Claim 14 (depends on 1)

The vector of claim 1, wherein the infectious (+) SS RNA virus is a nonpathogenic virus.

### Claim 15 (depends on 14)

The vector of claim 14, wherein the nonpathogenic virus is an attenuated virus.

### Claim 16 (depends on 1)

The vector of claim 1, wherein the promoter is a CMV promoter, a RSV promoter, a SV40 promoter, a HSV promoter, a human Pol I promoter, a human Pol II promoter, or a human Pol III promoter.

### Claim 17 (depends on 1)

A composition comprising the vector of claim 1 and a carrier.

### Claim 18 (depends on 14)

A pharmaceutical composition comprising the vector of claim 14 and a pharmaceutically acceptable carrier.

### Claim 19 (depends on 14)

A vaccine comprising a therapeutically effective amount of a vector of claim 14.

### Claim 20 (depends on 18)

A vaccine comprising a therapeutically effective amount of the pharmaceutical composition of claim 18.

### Claim 21 (depends on 1)

A homogenous population of clonally purified (+) SS RNA virus obtained from eukaryotic cells transfected with the vector of claim 1.

### Claim 22 (depends on 21)

A composition comprising the homogeneous population of clonally purified (+) SS RNA virus of claim 21 and a carrier.

### Claim 23 (depends on 21)

A pharmaceutical composition comprising the homogenous population of clonally purified (+)SS RNA virus of claim 21 and a pharmaceutically acceptable carrier.

### Claim 24 (depends on 14)

A vaccine comprising a therapeutically effective amount of a homogeneous clonally purified live (+)SS RNA virus population obtained from cells transfected with the vector of claim 14.

### Claim 25 (depends on 1)

A method of preparing a homogeneous clonally purified live (+)SS RNA virus population, wherein the method comprises transfecting the vector of claim 1 into a eukaryotic cell and isolating (+)SS RNA viruses, thereby obtaining the homogeneous clonally purified live (+)SS RNA virus population.

### Claim 26 (depends on 25)

The method of claim 25, wherein the eukaryotic cell is a Vero cell, a CHO cell, or a MDCK cell.

### Claim 27 (depends on 14)

A method of preparing a vaccine for protecting a subject against a disease caused by an infectious (+) SS RNA virus, wherein the method comprises transfecting the vector of claim 14 into a eukaryotic cell and isolating the (+) SS RNA viruses, thereby obtaining a vaccine.

### Claim 28 (depends on 27)

The method of claim 27, wherein the eukaryotic cell is a Vero cell, a CHO cell, or a MDCK cell.

### Claim 29 (depends on 20)

A method for protecting a subject against a disease caused by an infectious (+) SS RNA virus, wherein the method comprises administering the vaccine of claim 20 to a subject.

### Claim 30 (depends on 29)

The method of claim 29, wherein the subject is a mammal.

### Claim 31 (depends on 30)

The method of claim 30, wherein the mammal is a human or a veterinary animal.

### Claim 32

A method of preparing a stable plasmid, wherein the method comprises a DNA encoding a genomic RNA of an infectious (+) SS RNA virus, wherein the method comprises introducing at least three introns into the DNA, and wherein at least one intron is in the region encoding a structural protein of the (+) SS RNA virus and at least one intron is in the region encoding a non-structural protein of the (+) SS RNA virus.

### Claim 33 (depends on 1)

A method of preparing a vector of claim 1, wherein the method comprises transfecting the vector of claim 1 into a host cell and isolating the vector from the host cell.

### Claim 34 (depends on 1)

An isolated cell transfected with a vector of claim 1.

### Claim 35 (depends on 25)

A method of protecting a subject against a disease caused by an infectious (+) SS RNA virus, wherein the method comprises administering a vaccine comprising the homogeneous clonally purified live (+)SS RNA virus population of claim 25.