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Dengue virus E protein blocking peptide P4 and application thereof

A dengue virus and protein technology, applied in the direction of viral peptides, viruses, antiviral agents, etc., can solve the problem of no specific antiviral drugs, so as to inhibit dengue virus infection, reduce the infection rate, and reduce the risk of infection. effect of chance

Active Publication Date: 2016-07-20
CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the research on E protein mainly focuses on the research and development of vaccines. Dengue fever and severe dengue fever caused by dengue virus can only be treated clinically, and there is no specific antiviral drug.

Method used

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  • Dengue virus E protein blocking peptide P4 and application thereof
  • Dengue virus E protein blocking peptide P4 and application thereof
  • Dengue virus E protein blocking peptide P4 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Study on the Affinity of Small Molecular Peptide P4 and Dengue Virus Receptor Integrin β3

[0022] In this example, the SPR method was used to study the affinity between the small molecule peptide and the dengue virus receptor integrin β3. The small molecule peptide P4 was prepared into a solution with a certain concentration with phosphate buffer solution (PBS, pH7.4). Integrin ανβ3 (product number: RD3050-av-050) buffer was replaced with phosphate buffer and prepared to the desired concentration. The integrin ανβ3 was highly coupled to a CM5 chip (product number: GE29104988), and the small molecule peptide P4 was used as the mobile phase for affinity determination. BiacoreT200 was used for affinity analysis, and the results showed that the dissociation constant of small molecular peptide P4 and integrin β3 was 9.09E-5, indicating that it had a strong affinity with integrin β3 ( figure 1 ).

Embodiment 2

[0023] Example 2 Cytotoxicity Experiment of Small Molecular Peptide P4

[0024] In this example, MTT method was used to measure the cytotoxicity of small molecular peptides.

[0025] The small molecule peptide P4 was diluted with cell culture medium (DMEM containing 2% serum, 11965092, GIBCO) to prepare a solution with a certain concentration. MTT (3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenyltetrazolium romide) was purchased from Sigma (product number: SIGMAM5655). The cells used were HUVECs (Human Umbilical Vein Endothelial Cells).

[0026] The specific steps are as follows: when the HUVEC cells grow to 80%-90%, the original medium is discarded and replaced with 200 μl per well of maintenance solution added with drugs. Both drugs have 6 gradients, namely 5μM, 10μM, 20μM, 40μM, 80μM and 100μM, 37℃5%CO 2 Cultivate for 24h. After 24 hours, discard the maintenance solution containing the drug and replace it with a maintenance solution containing 10% MTT, 200 μl per well, at...

Embodiment 3

[0027] Example 3 Inhibitory Experiment of Small Molecular Peptide P4 on Dengue Virus Infection

[0028] The small molecule peptide P4 was diluted with cell culture medium (DMEM containing 2% serum, 11965092, GIBCO) to prepare a solution with a certain concentration. The cells used were HUVECs (Human Umbilical Vein Endothelial Cells).

[0029] The specific steps are as follows: HUVEC cells were inoculated into a 48-well plate, about 5 × 10 per well. 4 cells. 37°C 5% CO 2 Cultivate for 16h. Discard the medium, wash once with PBS, dilute small peptides with 1640 medium containing 2% FBS, and dilute to gradients of 40 μM, 20 μM, 10 μM, 5 μM, 2.5 μM, 1.25 μM and 0.3 μM, respectively. 100 μl per well, set blank control and positive control, 37°C 5% CO 2 Cultivate for 2h. After 2 hours, add 10 μl of virus solution (MOI=2) to each well, at 37°C in 5% CO 2 Culture for 1h. After 1 hour, the supernatant and cells were collected (3M glycine was used to remove extracellular virus),...

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Abstract

The invention relates to a dengue virus E protein blocking peptide P4 and an application thereof. The application comprises the following steps: performing bioinformatics analysis on the key E protein of the dengue virus infection host, synthesizing small molecular peptide P4 aiming at the key region ED-III design; combining a dynamics experiment and a cell experiment, researching the appetency between the small molecular peptide P4 and a receptor, and then researching the effect of the small molecular peptide P4 in blocking the virus to enter the host cell through the cellular experiment. The result shows that the small molecular peptide P4 and the receptor integrin beta3 have strong appetency, and can obviously inhibit the dengue virus to infect the host cell, thereby achieving the aim of blocking the dengue virus infection. The small molecular peptide P4 can be used for preparing the medicine and dengue vaccine to resist the dengue virus infection so as to effectively prevent and control the dengue virus.

Description

technical field [0001] The invention relates to the field of biomedical engineering, in particular to a dengue virus E protein blocking peptide P4 and its application. Background technique [0002] Dengue virus (DENV) is a single-stranded positive-sense RNA virus belonging to the family Flaviviridae, including four serotypes DENV1-4. The DENV genome is about 11kb, and an open reading frame encodes an envelope glycoprotein (E) Three structural proteins, membrane protein (M) and capsid protein (C), and seven nonstructural proteins NS1, NS2a, NS2b, NS3, NS4a, NS4b and NS5 (KuadkitkanA, WikanN, FongsaranC, SmithDR: Identification and characterization of prohibitina sareceptor protein mediating DENV-2entryintoinsect cells. Virology2010 , 406(1):149-161). The envelope protein E protein on the surface of the virus is the envelope glycoprotein and the largest structural protein on the DENV virion. It plays an important role in the process of virus adsorption, fusion with the host c...

Claims

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Application Information

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IPC IPC(8): C07K14/18A61K39/12A61P31/14
CPCA61K39/12C07K14/005C12N2770/24122C12N2770/24134Y02A50/30
Inventor 安静吴艳花崔小云范东瀛杨威
Owner CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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