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Flavivirus virus like particle

A yellow fever virus, virus-like technology, applied in the direction of viruses, viral peptides, antiviral agents, etc., can solve the problems of dengue fever limited to vector control measures, the theoretical risk of immune enhancement, and the lack of effective dengue fever treatment.

Active Publication Date: 2017-05-17
VLP THERAPEUTICS LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although dengue is the most important yellow fever virus in terms of global disease incidence, the development and use of vaccines against this virus has thus far been hampered by vaccine-associated adverse events, such as immune enhancement of infection and the need for induction of long-lasting protective immune responses with simultaneous against the theoretical risk of all four dengue serotypes)
[0005] There is no effective dengue treatment and prevention against dengue is currently limited to vector control measures

Method used

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  • Flavivirus virus like particle
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Preparation of virus-like particles comprising proteins or fragments of dengue virus type 1-2

[0085] Dengue virus type 1 and 2 virus structural proteins were used as wild-type virus structural proteins. At least one change is introduced in an envelope region of a viral structural protein. That is, amino acid Phe at position 108 of dengue virus type 1 envelope protein (SEQ ID NO: 20) becomes Ala (F108A), and amino acid Lys at position 203 of dengue virus type 1 envelope protein becomes Asn ( K203N) (envelope region: 172-676aa of SEQ ID NO: 4), the amino acid Asn at position 246 was changed to Met (K246M). In this example, the envelope region corresponds to 172-676aa of SEQ ID NO: 2, position 108 of SEQ ID NO: 20 corresponds to position 289 of SEQ ID NO: 2, and position 203 of SEQ ID NO: 20 corresponds to SEQ ID NO: Position 384 of ID NO: 2, and position 246 of SEQ ID NO: 20 corresponds to position 427 of SEQ ID NO: 2. In this example, mutations in this envelope re...

Embodiment 2

[0092] Preparation of virus-like particles containing dengue virus structural protein prM and envelope

[0093] The dengue virus structural protein expression vector containing prM and modified envelope protein F108A, K203N or K203N+F108A (respectively SEQ ID NO1, 3, 15) used in Example 1 was used. In the same manner as in Example 1, 20ug of the vector was transfected into 293F cells and cultured. On day 4 post-transfection, supernatants were harvested from transfected cell cultures. Western blotting was performed to detect DENV VLPs using a monoclonal antibody (9.F.10, Santa Cruz Biotech) as the primary antibody and a rabbit anti-mouse IgG-HRP conjugated antibody as the secondary antibody. The results are shown in Figure 4 . as by Figure 4 It was shown that cells transfected with DENV1-prM and envelope proteins with at least one change produced higher amounts of envelope proteins.

[0094] Filter the supernatant using a 0.45 μm filter to obtain virus-like particles. ...

Embodiment 3

[0096] Antibodies against dengue virus

[0097] The purified VLP (K203N+F108A) obtained in Example 2 was named DEN1 VLP and used in this example. The purified virus-like particles were further concentrated using a spin column (molecular weight cutoff: 100 kDa) to prepare virus-like particles for immunization. Then, 4 mice were immunized with DEN1 VLP 30 μg in PBS containing aluminum adjuvant (alhydrogel 2%, Sergeant Aduvants) by intramuscular injection twice at 0 and 4 weeks.

[0098] Dengue virus-specific IgG titers of sera derived from immunized mice were determined by ELISA system. Neutralizing antibodies against each of DENV-1, DENV-2, DENV-3 and DENV-4 viruses in the sera were determined. The following dengue serotypes 1 to 4 were used:

[0099]DENV-1, Philippine-99St12A strain

[0100] DENV-2, Philippine-00St22A strain

[0101] DENV-3, Philipine-SLMC50 strain

[0102] DENV-4, Philippine-SLMC318 strain

[0103] JEV-JaOAr S-982 strain.

[0104] The anti-DENV neut...

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Abstract

The present application provides a virus like particle comprising one or more flavivirus structural proteins, and a composition or vaccine comprising thereof, its use in the prevention or treatment of flavivirus infection. The flavivirus structural protein contains at least one amino acid alteration in the envelope region. Examples of flavivirus contains dengue virus.

Description

technical field [0001] The present application relates to virus-like particles comprising one or more yellow fever virus structural proteins, as well as compositions or vaccines comprising them, for their use in medicine, especially in the prevention or treatment of yellow fever virus infection. Background technique [0002] Yellow fever virus comprises more than 70 different viruses, many of which are arthropod vectors and are transmitted by mosquitoes or ticks. [0003] Yellow fever virus is a genus of viruses in the Flaviviridae family. The genus includes West Nile virus (WNV), dengue virus (DENV), tick-borne encephalitis virus (TBEV), yellow fever virus (YFV), Japanese encephalitis virus (JEV), hepatitis C virus (HCV), and several other viruses that may cause encephalitis or hemorrhagic disease. [0004] Dengue fever is a mosquito-borne disease caused by the yellow fever virus and has spread to most tropical and many subtropical regions. The disease is caused by four ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C07K19/00
CPCA61K2039/5258A61K2039/575A61K2039/53C12N2770/24123C12N2770/24134A61K39/12C07K14/005C12N7/00C12N2770/24023A61P31/14Y02A50/30
Inventor 赤畑涉上野隆司
Owner VLP THERAPEUTICS LLC
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