US-12622958-B2 - Virus-like particles containing RSV antigen protein and vaccines using the same

US12622958B2US 12622958 B2US12622958 B2US 12622958B2US-12622958-B2

Abstract

An RSV virus-like particle which includes a chimeric protein that includes a core consisting of an influenza M1 protein, an RSV-derived preF protein, an RSV-derived G protein or part of G protein displayed on the surface of the core, can exhibit excellent effects in terms of inhibiting RSV virus infection and inhibiting the inflammatory response of the lungs.

Inventors

Fu-Shi Quan
Ki-Back Chu
Su-Hwa Lee

Assignees

UNIVERSITY-INDUSTRY COOPERATION GROUP OF KYUNG HEE UNIVERSITY

Dates

Publication Date: 20260512
Application Date: 20221103
Priority Date: 20211214

Claims (6)

1 . A respiratory syncytial virus (RSV)-like particle comprising: a core consisting of an influenza virus matrix protein 1 (M1); and an antigenic protein displayed on the surface of the core, wherein the antigen protein comprises a preF protein derived from RSV consisting of an amino acid sequence of SEQ ID NO: 12, and a chimeric protein, which comprises a first tandem sequence consisting of an amino acid sequence of SEQ ID NO: 9 and a second tandem sequence consisting of an amino acid sequence of SEQ ID NO: 10.
2 . The RSV virus-like particle of claim 1 , wherein the antigen protein further comprises: an RSV-derived G protein consisting of an amino acid sequence of SEQ ID NO: 8.
3 . The RSV virus-like particle of claim 2 , wherein the chimeric protein is one in which the transmembrane domain and cytoplasmic tail domain of influenza hemagglutinin are linked to the C-terminus of the second tandem sequence.
4 . The RSV virus-like particle of claim 2 , wherein the chimeric protein is one in which the first tandem sequence and the second tandem sequence are linked by a linker.
5 . A composition that induces an immune response against RSV infection comprising the RSV virus-like particle of claim 1 as an active ingredient.
6 . The composition of claim 5 , wherein the composition further comprises an adjuvant.

Description

TECHNICAL FIELD This application claims priority to Korean Patent Application No. 10-2021-0178199, filed on Dec. 4, 2021. The present disclosure relates to a virus-like particle including an RSV antigen protein and a vaccine using the same. REFERENCE TO AN ELECTRONIC SEQUENCE LISTING The contents of the electronic sequence listing (2022-11-03_SequenceListing.xml; Size: 19,458 bytes; and Date of Creation: Nov. 3, 2022) is herein incorporated by reference in its entirety. BACKGROUND The scientific name of respiratory syncytial virus (RSV) is Human orthopneumovirus. RSV is a negative-sense single-stranded RNA virus with a lipid envelope. In the lipid envelope of RSV, there are G proteins and F proteins, which are surface proteins. G proteins and F proteins are the two major surface proteins involved in viral attachment and early stages of infection, and these proteins are major targets of antibodies. The G protein is a glycoprotein and plays a role in attaching the virus to the ciliary cells of the host airway. The F protein is an abbreviation of a fusion protein and it serves to fuse the cell membranes of a virus and a host to form a syncytium. The F protein has several forms. Before binding to a host cell, the F protein is called preF (prefusion state). The preF protein exists in a trimeric form and includes an antigenic site Ψ. When RSV binds to a host cell, the preF loses Ψ, undergoes a change in its shape, is inserted into the host cell membrane, and induces a fusion of the virus with the host cell membrane. The F protein which has undergone a conformational change becomes stable and is in a long postfusion state (PostF). To date, there is no licensed vaccine to prevent RSV infection. The RSV vaccines reported to the academic community are not highly immunogenic and have a side effect that induces an inflammatory response in the lungs; therefore, they have not been commercialized. Prior Document Patent Document (Patent Document 1) Korean Patent Application Publication No. 10-1862137 (May 1, 2017) Non-Patent Document (Non-Patent Document 1) Virus-like particle vaccines expressing Toxoplasma gondii rhoptry protein 18 and microneme protein 8 provide enhanced protection. Vaccine 2018, 36, 5692-5700 SUMMARY According to a specific embodiment, the present disclosure provides a virus-like particle (VLP), which includes influenza M1, RSV-derived preF, and chimeric protein comprising all or some residue of RSV-derived G proteins, and an RSV vaccine using the same. An aspect of the present disclosure provides a respiratory syncytial virus (RSV)-like particle, which includes: a core consisting of an influenza virus matrix protein 1 (M1); and an antigenic protein displayed on the surface of the core, wherein the antigen protein includes a preF protein derived from RSV consisting of an amino acid sequence of SEQ ID NO: 12. The RSV-derived preF protein is a codon-optimized sequence based on the sequence of GenBank: MN125707.1, and the immunization effect can be significantly enhanced when used as a VLP in combination with M1 than when used alone. In a specific embodiment, the antigen protein may further include an RSV-derived G protein; or a chimeric protein, which includes a first tandem sequence consisting of an amino acid sequence of SEQ ID NO: 9 and a second tandem sequence consisting of an amino acid sequence of SEQ ID NO: 10. The present inventors prepared various VLPs by combining RSV-M or influenza M1 as a core and RSV-preF, RSV-G, and RSV-Gt as an antigen protein; infected VLP-immunized mice with RSV to determine the viral titer and the degree of pulmonary inflammatory response. As a result, they found that the VLPs in which M1 core protein and antigenic protein were combined increased the immunization effect against RSV and reduced the inflammatory side effects of the lungs, that the immunization effect of VLPs in which RSV-preF was combined with M1 was significantly superior to that of VLPs in which RSV-preF was used alone, and that the effect was further enhanced when RSV-preF and RSV-G or Gt were combined. Additionally, the present inventors found that in the process of producing a chimeric protein including a part of the nucleotide sequence of G protein, if the 151st to the 261st and the 151st to the 260th nucleotides of the nucleotide sequence encoding the G protein are used, the expression did not go smoothly, whereas when the nucleotides from the 150th to the 260th of the nucleotide sequence were used, the expression went smoothly. Additionally, the protein expressed from the nucleotide sequence from the 150th to the 260th nucleotides consists of an amino acid sequence different from the original G protein due to changes in triplet codons, and that it has an excellent immunization effect of the modified protein. The virus-like particle (VLP) is a non-infectious particle because it includes a protein of a virus but does not include a genetic material. Virus-like particles can be synthesized by individua