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Polypeptide and composition for restraining SARS-COV-2 infection, and purposes of polypeptide and composition

A technology of SARS-COV-2 and a composition, applied in the field of biomedicine, can solve the problems of difficult prevention and treatment of SARS-COV-2, and achieve the effects of inhibiting virus invasion, safe use, and no toxic and side effects

Active Publication Date: 2020-07-31
北京中科微盾生物科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the difficult problem of prevention and treatment of SARS-COV-2, the present invention provides a polypeptide that inhibits SARS-COV-2, which can specifically bind to the RBD region of the S protein of SARS-COV-2, preventing Break the combination of virus and cell receptors, and then effectively inhibit the infection of the virus from the source

Method used

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  • Polypeptide and composition for restraining SARS-COV-2 infection, and purposes of polypeptide and composition
  • Polypeptide and composition for restraining SARS-COV-2 infection, and purposes of polypeptide and composition
  • Polypeptide and composition for restraining SARS-COV-2 infection, and purposes of polypeptide and composition

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] 1. Fake virus packaging

[0062] 1.1 Plasmid extraction

[0063] Pseudovirus system plasmid construction, constructing the S protein of full-length SARS-COV-2 into the multiple cloning site of PSV (pcDNA3.1-SARS-COV-2-S, wherein the S protein removes 19 amino acids at the C-terminal), Transform DH5-a competent cells simultaneously with the backbone plasmid pNL-LUC-E- (this plasmid is a universal pseudovirus packaging plasmid). After heat shock at 42°C for 90 seconds, coat LB plates with an ampicillin concentration of 5 micrograms per milliliter. Incubate overnight in a 37°C incubator.

[0064] PSV map such as Figure 5 shown.

[0065] Pick a single colony and inoculate LB liquid medium with an ampicillin concentration of 5 micrograms per milliliter. Shake the bacteria overnight at 37°C.

[0066] Centrifuge at 12000 for 10 minutes, collect the bacteria, and extract the plasmid with Tiangen Plasmid Extraction Kit. Nanodrop measures DNA concentration.

[0067] 1.2 ...

Embodiment 2

[0105] Verification of the inhibitory effect of polypeptide, fucoidan and composition on pseudovirus infection:

[0106] The SARS-COV-2 pseudovirus was incubated with 91.06 μM / L fucoidan and 10 μM / L VDP-4 alone, or the virus was co-incubated with 91.06 μM / L fucoidan and 10 mM / L VDP-4, and set up a control group. After mixing and incubating the virus with the above components, add it to Vero-E6 cells, incubate on ice for 45 minutes, wash the cells twice with serum-free DMEM, lyse the cells with Trizol reagent, extract total RNA, and analyze the copy number of the pseudovirus. The results are as follows: figure 2 shown.

[0107] Depend on figure 2 It can be seen that:

[0108] After VDP-4 was mixed with virus alone, the virus-cell binding efficiency was reduced to 80%; after fucoidan was mixed with virus alone, the virus-cell binding efficiency was reduced to 77%; VDP-4 and fucoidan co-existed After the composition is mixed with the virus and incubated, the virus-cell bind...

Embodiment 3

[0111] The effect of VDP-4 mixed with fucoidan (91.06μM / L) at different concentrations to inhibit the invasion of SARS-COV-2 pseudovirus.

[0112] Mix and incubate the SARS-COV-2 pseudovirus with the above-mentioned mixture of different concentrations (diluted in DMEM) at a dose of MOI=10, add the incubated virus into the cells, infect for one hour, discard the supernatant, and wash twice with DMEM , add serum-containing medium, and cultivate at 37 degrees. Another group of cells was directly incubated with the above-mentioned mixtures of different concentrations for 1 hour, the supernatant was discarded, washed twice with DMEM, infected with virus at MOI=10, and luciferase reporter gene detection was carried out 48 hours later. Test results such as image 3 shown.

[0113] Depend on image 3 It can be seen that the SARS-COV-2 virus and the composition treat the cells separately, and the virus infection will not be significantly affected. However, mixing the SARS-COV-2 vir...

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Abstract

The invention provides polypeptide for restraining SARS-COV-2 infection, and belongs to the field of biopharmaceuticals. The polypeptide is VDP-4, and has an amino acid sequence as shown in SEQID NO:1; and the polypeptide can also be a VDP-4 modifying product, or one of pharmaceutically acceptable salts, esters and prodrugs. The polypeptide can be specifically bound with an RBD region of an S protein of SARS-COV-2 to block combination of viruses and cell receptors and further effectively restrain virus infection from the source. The invention also provides a composition for restraining the SARS-COV-2, and a purpose of the composition.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a polypeptide and a composition for inhibiting SARS-COV-2 infection, and the use of the polypeptide and the composition in preparing medicines for preventing and / or treating SARS-COV-2 infection. Background technique [0002] Since the end of 2019, the SARS-COV-2 epidemic has broken out successively around the world. Since the virus is a brand new virus and is mainly transmitted through droplets and contact, the virus spread rapidly in a short period of time, causing a huge impact on the world. Since the vaccine against SARS-COV-2 is still in the research stage, it is difficult to apply it on a large scale in the short term, and its protective efficiency and safety need to be verified through a large number of experiments. In addition, reasons such as virus mutation may further increase the difficulty of preventing and treating SARS-COV-2. Therefore, there is an urgent need to develop n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P31/14A61K31/715
CPCA61K31/715A61K38/00A61P31/14C07K7/08A61K2300/00
Inventor 岳少恒任金成
Owner 北京中科微盾生物科技有限责任公司
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