Search

KR-102962029-B1 - Recombinant bacteriophage nanofiber for preventing Enterovirus 71 infection and a composition comprising thereof

KR102962029B1KR 102962029 B1KR102962029 B1KR 102962029B1KR-102962029-B1

Abstract

The present invention relates to a bacteriophage expressing an epitope containing the amino acid sequence of KQEKD and a composition for preventing enterovirus 71 infection containing the same. The vaccine using the recombinant bacteriophage according to the present invention is safer to use than live vaccines or attenuated live vaccines, and has superior immune response induction ability compared to peptide vaccines, so it has the advantage of being usable for the prevention of hand, foot, and mouth disease for which no preventive vaccine or treatment has yet been developed.

Inventors

  • 진효언
  • 장선영

Assignees

  • 아주대학교산학협력단

Dates

Publication Date
20260508
Application Date
20230410
Priority Date
20220411

Claims (9)

  1. A recombinant bacteriophage in which an epitope containing the amino acid sequence of KQEKD (SEQ No. 1) is expressed, said epitope is expressed to be displayed on the p8 major coat protein of bacteriophage f88.
  2. delete
  3. delete
  4. A vaccine composition comprising the recombinant bacteriophage of claim 1.
  5. In paragraph 4, the vaccine composition is a vaccine composition for preventing enterovirus 71 infection.
  6. In paragraph 4, the vaccine composition is administered via a route selected from the group consisting of oral administration, intravenous administration, intradermal administration, intraperitoneal administration, subcutaneous administration, and intramuscular administration.
  7. In paragraph 4, the vaccine composition is administered 1 to 5 times.
  8. A composition for preventing hand, foot, and mouth disease comprising the recombinant bacteriophage of claim 1.
  9. A composition for preventing hand, foot, and mouth disease according to claim 8, wherein the hand, foot, and mouth disease is caused by an enterovirus 71 infection.

Description

Composition for preventing Enterovirus 71 infection comprising recombinant bacteriophage {Recombinant bacteriophage nanofiber for preventing Enterovirus 71 infection and a composition comprising thereof} The present invention relates to a composition for preventing enterovirus 71 infection comprising a recombinant bacteriophage, and more specifically, to a bacteriophage expressing an epitope comprising the amino acid sequence of KQEKD and a composition for preventing enterovirus 71 infection comprising the same. Enterovirus 71 is a virus that causes hand, foot, and mouth disease when it infects infants and young children, and it can leave more severe aftereffects compared to Coxsackievirus, another virus that causes hand, foot, and mouth disease. Enterovirus 71, or EV71 (BrCr lineage), was first isolated and identified in the United States in 1969, and following small-scale outbreaks in Japan and Sweden in 1973, successive waves of EV71 outbreaks have been reported globally in the United Kingdom, Australia, Sweden, Bulgaria, Japan, China, Hong Kong, Taiwan, Malaysia, and Singapore. In particular, over the past decade, large and small-scale outbreaks associated with neurological complications and deaths, such as aseptic meningitis, fatal encephalitis, and polio-like paralysis, have been reported in the Asia-Pacific region. EV71, which emerged in Taiwan in 1998, infected thousands of children and resulted in 405 cases of severe neurological disease and 78 child deaths, while in China in 2008, hand, foot, and mouth disease (HFMD) caused by enterovirus EV71 resulted in approximately 3.4 million cumulative cases and 1,400 deaths. As such, EV71 has established itself as a major neurotoxic virus since polio was nearly completely eradicated. EV71, a typical member of the Picornaviridae family, is a small, envelope-less, positive-stranded RNA virus containing four capsid proteins: VP1, VP2, VP3, and VP4. These capsid proteins form an icosahedral structure in which VP1-3 are exposed on the surface of the virus and VP4 is arranged internally. Among these capsid proteins, VP1 is considered a major cause of viral pathogenesis, playing a crucial role in the processes of adsorption and shedding during viral infection (The efficacy of viral capsid inhibitors in human enterovirus infection and associated diseases. Li C. et al., (2007) Curr Med Chem 14: 847-856). Meanwhile, bacteriophages were discovered by Fredrick Twart (1915) and Felix d'Herelle (1917). Bacteriophages are viruses that possess a DNA or RNA genome and infect bacteria to replicate within them. Bacteriophages can undergo lytic or lysogenic cycles within bacteria. During a lytic cycle, the genetic material of the bacteriophage is injected into the bacteria, where transcription, translation, and replication occur. This leads to the assembly and packaging of bacteriophage proteins and nucleic acids, ultimately resulting in lysis. At this point, a large number of bacteriophages are released, allowing them to easily infect additional bacteria. Some bacteriophages can also undergo a lysogenic cycle, during which the bacteriophage genetic material binds to the bacterial genome. Recently, genetically modified bacteriophages are being developed as vaccines or for targeted delivery to kill cancer cells (Therapeutic and prophylactic applications of bacteriophage components in modern medicine. Adhya S et al. Cold Spring Harb Perspect Med. (2014) 1, 1; Killing cancer cells by targeted drug-carrying phage nanomedicines Bar H. et al. BMC Biotechnol.(2008) 37; Phage protein-targeted cancer nanomedicines, Petrenko VA and Jayanna PK. FEBS Lett. (2014) 588:341). This is based on bacteriophage display technology, specifically, to use a bacteriophage as a carrier, an exogenous gene is inserted into the bacteriophage genome, and the exogenous gene is exposed on the surface of the virus during the self-assembly process of the bacteriophage along with the expression of the bacteriophage genome, so that the epitope is displayed on the surface of the bacteriophage in the form of a fusion protein as part of the envelope protein. The inventors completed the present invention by confirming that, while seeking a method to prevent infection with hand, foot, and mouth disease for which no vaccine or treatment has yet been developed, a recombinant bacteriophage having an epitope containing the amino acid sequence of KQEKD expressed on the surface of the bacteriophage can effectively produce VP1 antigen-specific antibodies, and that these antibodies can effectively prevent enterovirus 71 infection. Figure 1 shows an AFM image of a recombinant f88-K7G phage according to one embodiment of the present invention. Figure 2 is the result of confirming epitope peptide expression by recombinant f88-K7G phage according to one embodiment of the present invention using SDS-PAGE. Figure 3 is the result of confirming epitope peptide expression by recombinant f88-K7G phage according to one embodiment of th