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DENV (Dengue virus) protein E blocking peptide P7 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, achieve inhibition of dengue virus infection, no toxic side effects, and reduce infection rates Effect

Active Publication Date: 2016-07-27
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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  • DENV (Dengue virus) protein E blocking peptide P7 and application thereof
  • DENV (Dengue virus) protein E blocking peptide P7 and application thereof
  • DENV (Dengue virus) protein E blocking peptide P7 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Study on Affinity of Small Molecular Peptide P7 and Dengue Virus Receptor Integrin β3

[0024] 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 P7 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 P7 was used as the mobile phase for affinity determination. BiacoreT200 was used for affinity analysis, and the results showed that the dissociation constants of the small molecular peptide P7 and integrin β3 were 7.93E-5, indicating that it has a strong affinity with integrin β3 ( figure 1 ). Example 2 Cytotoxicity Experiment of Small Molecular Peptide P7 ...

Embodiment 2

[0024] 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 P7 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 P7 was used as the mobile phase for affinity determination. BiacoreT200 was used for affinity analysis, and the results showed that the dissociation constants of the small molecular peptide P7 and integrin β3 were 7.93E-5, indicating that it has a strong affinity with integrin β3 ( figure 1 ). Example 2 Cytotoxicity Experiment of Small Molecular Peptide P7

[0025] In this example, MTT method was used to measure the cytotoxicity of small molecular peptides.

...

Embodiment 3

[0028] Example 3 Inhibitory Experiment of Small Molecular Peptide P7 on Dengue Virus Infection

[0029] The small molecule peptide P7 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).

[0030] 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 0.325 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 DENV (Dengue virus) protein E blocking peptide P7 and an application thereof. According to the invention, key protein E of a DENV infected host is subjected to bioinformatic analysis, a small molecule peptide P7 is designed and synthesized by aiming at a critical area ED-III of the protein E, affinity between the peptide and a receptor is researched through combination of dynamic experiments and cell experiments, and the effect of the peptide for blocking the virus from entering host cells is researched through cytology experiments. A result indicates that the small molecule peptide P7 has higher affinity with the receptor integrin beta 3 and can remarkably inhibit the DENV from infecting the host cells, thereby blocking the DENV infection. Drugs and dengue vaccines resistant to DENV infection are prepared from the small molecule peptide P7, and accordingly, the DENV can be effectively prevented and treated.

Description

technical field [0001] The invention relates to the field of biomedical engineering, in particular to a dengue virus E protein blocking peptide P7 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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