Polypeptide capable of inhibiting infection of Zika virus, dengue virus and yellow fevervirus and application of polypeptide

A Zika virus and polypeptide technology, applied in the field of polypeptide virus inhibitors, can solve the problems of loss of infectivity, genome loss, and destruction of virus envelope structure, and achieve good antiviral effect and safety in vivo, broad-spectrum anti-jaundice The effect of viruses

Active Publication Date: 2020-06-23
FUDAN UNIV
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

Some studies have found that the polypeptide Z2 from the stem region of the Zika virus E protein can inhibit the infection of the Zika virus, but its mechanism of action is mainly to destroy the viral envelope structure, resulting in the loss of the genome and thus loss of infectivity. In essence, it does not block fusion fusion inhibitor

Method used

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  • Polypeptide capable of inhibiting infection of Zika virus, dengue virus and yellow fevervirus and application of polypeptide
  • Polypeptide capable of inhibiting infection of Zika virus, dengue virus and yellow fevervirus and application of polypeptide
  • Polypeptide capable of inhibiting infection of Zika virus, dengue virus and yellow fevervirus and application of polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1 polypeptide drug design

[0040] In this example, multiple polypeptides were designed based on the amino acid sequence of the ZIKV envelope protein (Envelope, E), and were screened by plaque reduction experiments. After screening, a polypeptide with good anti-Zika virus effect was obtained, and the polypeptide was named It is Z155, the amino acid sequence of which is shown in Z155-SEQ1; the sequence is: FSQILIGTLLMWLGL.

Embodiment 2

[0041] Example 2 Virus plaque assay detects the inhibitory activity of polypeptide Z155 on ZIKV infection

[0042] (1) Take well-grown BHK-21 cells and spread them on a 24-well plate, 5×10 cells per well 4 cells, cultured overnight for 16-24 hours;

[0043] (2) Dissolve the polypeptide in DMSO after weighing, measure the OD280 absorbance, and calculate the polypeptide concentration according to the extinction coefficient;

[0044] (3) Using serum-free DMEM starting from an initial concentration of 10 μM, dilute polypeptide Z155 in a 2-fold ratio in EP tubes, with a volume of drug-containing medium in each well of 160 μL, and dilute a total of 6 gradients, each gradient setting 3 duplicate holes. Set random peptide controls;

[0045] (4) Use serum-free DMEM to dilute ZIKV so that the final infection concentration of the virus is 0.25×10 3 For each ml of PFU, add 160 μL of virus dilution solution to each tube in (2) (to ensure that 25-35 plaques are formed in each well), and...

Embodiment 3

[0051] Example 3 Polypeptide Z155 inhibits the stage detection of ZIKV infection

[0052] (1) Spread BHK-21 cells in a 24-well plate, 5×10 per well 4 at 37°C, 5% CO 2 Cultivate under conditions for 12-16h;

[0053] (2) Adding the polypeptide Z155 at different time points, and different treatments after adding the polypeptide:

[0054] 1) Time-of-addition assay: After the virus and cells were incubated for different times (0h, 1h, 2h, 3h), 300μl of polypeptide Z155 was added, and the time points were respectively recorded as 0h, 1h, 2h, 3h post-infection. Record it as the drug well, and set the virus-infected positive control group and the cell well as the negative control at the same time, set 3 replicates for each group, the final concentration of the polypeptide is 5 μM, and the virus amount is 40-50 PFU per well;

[0055] 2) Cell wash assay: Add 300 μl of polypeptide Z155 with a concentration of 5 μM to the cells, incubate at 4°C for 30 minutes, absorb the supernatant, a...

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Abstract

The invention belongs to the technical field of biotechnology and biological medicines, relates to a virus inhibitor, and in particular to a polypeptide Z155 and an application thereof in preparing apolypeptide inhibitor for single infection or co-infection of Zika virus (ZIKV), dengue virus (DENV) and yellow fevervirus (YFV). Experiments show that the polypeptide Z155 can be combined with Zika virus particles and further inhibits virus infection cells through combinination with virus membrane-melting peptides. The polypeptide has a broad-spectrum anti-virus effect on flavivirus, and the cytotoxicity of the polypeptide is extremely low. The polypeptide inhibitor can be prepared into a medicament for treating ZIKV infection, is beneficial to treatment of single infection or co-infection ofZIKV, DENV and YFV, and is particularly important to treatment of Zaika infected persons, particularly infected pregnant women.

Description

technical field [0001] The invention belongs to the technical fields of biotechnology and biomedicine, and relates to polypeptide virus inhibitors. In particular, it relates to an inhibitor for inhibiting Zika virus (Zikavirus, ZIKV), dengue virus (dengue virus, DENV) and yellow fever virus (yellowfevervirus, YFV) infection, especially relates to polypeptide Z155 and its preparation of ZIKV, DENV and YFV infection Uses in polypeptide inhibitors. Background technique [0002] The prior art discloses that Zika virus (ZIKV) is a newly discovered arbovirus (arboviruses), which belongs to the genus Flavivirus of the family Flaviviridae. Zika virus is mainly transmitted through mosquito bites and has a natural foci. From 2013 to 2014, ZIKV broke out in French Polynesia. In April 2015, ZIKV infection broke out in Brazil. The virus spread rapidly in the Americas, and then ZIKV infection appeared in more than 60 countries around the world. From 2015 to January 2018, more than 800,0...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C12N15/40A61K38/10A61P31/14
CPCC07K7/08A61P31/14A61K38/00Y02A50/30
Inventor 陆路姜世勃孟余司露露
Owner FUDAN UNIV
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