# US 10,369,209 B2 · Drug target and construct for Japanese encephalitis virus infection

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- Number: US 10,369,209 B2
- Document ID: US10369209B2
- Status: Granted
- Kind: B2
- Application number: 15/303,375
- Filed: 2015-04-10
- Published: 2017-07-13
- Granted: 2019-08-06
- Earliest claimed: 2014-04-11
- Assignees: Utah State University
- Applicants: Young-Min Lee
- Inventors: Young-Min Lee
- Web page: https://patentgrep.com/patent/US10369209B2

## CPC

- A61K 39/12: Viral antigens
- 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)
- C12Q 1/18: Testing for antimicrobial activity of a material
- G01N 33/502: {for testing non-proliferative effects}
- A61K 2039/525: {Virus}
- C12N 2770/24111: Flavivirus, e.g. yellow fever virus, dengue, JEV
- 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 2770/24143: viral genome or elements thereof as genetic vector
- C12N 2770/24152: relating to complementing cells and packaging systems for producing virus or viral particles
- G01N 2333/18: Togaviridae; Flaviviridae
- G01N 2333/185: Flaviviruses or Group B arboviruses, e.g. yellow fever virus, japanese encephalitis, tick-borne encephalitis, dengue
- G01N 2800/26: Infectious diseases, e.g. generalised sepsis
- 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)
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

A group of mosquito-borne flaviviruses that cause fatal encephalitis in humans is among the most important of all emerging human pathogens of global significance. This group includes Japanese encephalitis virus (JEV), West Nile virus, St. Louis encephalitis virus, and Murray Valley encephalitis virus. In the present disclosure, the first reverse genetics system has been developed for SA14-14-2, a live JE vaccine that is most commonly used in most JE-endemic areas, by constructing an infectious bacterial artificial chromosome that contains the full-length SA14-14-2 cDNA. Using this infectious SA 14-14-2 cDNA, combined with a mouse model for JEV infection, a key viral neurovirulence factor has been discovered that is a conserved single amino acid in the ij hairpin adjacent to the fusion loop of the viral E glycoprotein, which regulates viral infectivity into neurons within the central nervous system.

## Claims

### Claim 1

A DNA construct comprising a cDNA sequence that encodes a JEV genomic RNA and has a nucleotide sequence set forth in SEQ ID NO:1 or SEQ ID NO:32;
wherein the DNA construct comprises a nucleotide sequence set forth in SEQ ID NO:2 when the cDNA sequence has the nucleotide sequence set forth in SEQ ID NO:1, and wherein the DNA construct comprises a nucleotide sequence set forth in SEQ ID NO:33 when the cDNA sequence has the nucleotide sequence set forth in SEQ ID NO:32.

### Claim 2 (depends on 1)

The DNA construct of claim 1, further comprising a bacterial artificial chromosome.

### Claim 3 (depends on 1)

The DNA construct of claim 1, wherein the DNA construct consists of the nucleotide sequence set forth in SEQ ID NO:2 or SEQ ID NO:33.

### Claim 4

A DNA construct comprising a nucleic acid sequence that encodes a JEV genomic RNA and has at least about 70% identity to a nucleotide sequence as set forth in SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17;
wherein the nucleic acid sequence is the nucleotide sequence as set forth in SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or SEQ ID NO:17.

### Claim 5 (depends on 1)

A vector comprising the DNA construct of claim 1.