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Application of salinomycin in preparation of anti-coronavirus drugs

A coronavirus and salinomycin technology, applied in the field of biomedicine, can solve problems such as the shortage of anti-coronavirus drugs, achieve the effects of shortening the research and development cycle, safety and reliability, and saving development costs

Active Publication Date: 2021-05-28
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Based on the pAPN receptor, the inventors have screened out compounds that are effective in inhibiting coronaviruses, mainly TGEV, aiming to provide a potential drug for treating TGE in clinical production, so as to overcome the shortage of anti-coronavirus drugs in the prior art

Method used

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  • Application of salinomycin in preparation of anti-coronavirus drugs
  • Application of salinomycin in preparation of anti-coronavirus drugs
  • Application of salinomycin in preparation of anti-coronavirus drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Computer-aided drug design screening of small molecule inhibitors based on pAPN

[0034] 1 Experimental method:

[0035] 1.1 SYBYL-X 2.1.1 Screening compounds

[0036] According to the Basics Manual SYBYL-X manual, the pAPN crystal structure (4fke) downloaded from the PDB website and the small molecule compound library were prepared before docking, and then docking screening was performed in Screen and GeomX modes to screen out the top ranked small molecules compound.

[0037] 1.2 PyMOL 2.4 Simulation binding

[0038] The compound salinomycin with the highest score above was used for binding simulation to find the amino acid site where pAPN binds to salinomycin. The binding hydrogen bond is marked with a dotted line. The 5A area near the pocket is presented in the form of lines, and other parts are presented in the form of cartoons.

[0039] like figure 1 As shown, the results show that the combination score of salinomycin and pAPN is 8.78, indicating tha...

Embodiment 2

[0041] Example 2 The toxic effect of salinomycin on PK-15 cells

[0042] 1 Experimental method

[0043] 1.1 Culture of wild-type PK-15 cells

[0044] Digest the PK-15 cells in the logarithmic growth phase from a 10cm-diameter culture dish, and dilute to 10% with DMEM complete medium containing 10% fetal bovine serum according to the cell density. 6cells / mL cell suspension, mix the cells evenly and inoculate them into a 96-well plate with a row gun, add 100 μL of cell suspension to each well, add the same volume of PBS to the surrounding wells, and gently tap around the culture plate to make The cells are evenly distributed, and after the cells have settled to the bottom of the culture plate, put them in 37°C, 5% CO 2 cultured in a cell culture incubator.

[0045] When the cells grow to 70%-80%, remove the medium, wash the cells once with sterile PBS, then add serum-free DMEM medium, and remove the medium after 12 hours after the cell cycle is synchronized.

[0046] 1.2 Cel...

Embodiment 3

[0053] Example 3 Salinomycin for the inhibitory effect of TGEV in PK-15

[0054] 1 Virus inoculation and salinomycin treatment

[0055] Press PK-15 cells by 10 6 Add one per well into a 6-well cell culture plate, wait for the cells to grow to 70-80%, inoculate TGEV virus fluid, and observe the cytopathic effect (cytopathic effect, CPE) every day.

[0056] One hour after virus inoculation, salinomycin was added to each well to a final concentration of 5 μM and 15 μM, observed after 24 hours and 36 hours, and the cell supernatant and cells were collected respectively.

[0057] 2 The effect of salinomycin on inhibiting the copy number of TEGV

[0058] RNA was extracted from the cell supernatant, reverse-transcribed into cDNA (RR047A, Takara), and quantified by fluorescence (RR820A, Takara). The reaction conditions were as follows: initial template denaturation at 95°C for 1 min; denaturation at 95°C for 20s; annealing at 58°C , 30s; extension at 68°C, 30s; 35 cycles, all tests...

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Abstract

The invention discloses application of salinomycin in preparation of an anti-coronavirus drug, and belongs to the technical field of biological medicines. Specifically, the invention discloses application of salinomycin in preparation of the anti-TGEV medicine, and tests prove that salinomycin with the concentration of 0.2-5 mu M has no obvious toxic effect on porcine kidney cells PK-15, salinomycin with the concentration of 5 mu M can significantly inhibit the copy number of TGEV in infection of PK-15, and salinomycin can become a new choice for preparation of the anti-TGEV medicine. The salinomycin is newly used as an old medicine, pharmacokinetic data of salinomycin on the market are detailed, the safety is reliable, second-stage clinical evaluation can be carried out quickly through development of the new application, the research and development period is shortened, the development cost is saved, and resources can be utilized to the maximum extent.

Description

technical field [0001] The invention relates to the technical field of biomedicine, and relates to the application of salinomycin in the preparation of anti-coronavirus drugs, in particular to the application of salinomycin in the preparation of anti-coronavirus TGEV drugs. Background technique [0002] Porcine transmissible gastroenteritis (TGE) is an acute, highly contagious gastrointestinal infectious disease caused by porcine transmissible gastroenteritis virus (TGEV), which was first reported in the United States in 1945 and subsequently spread to the world everywhere. With the rapid development of the pig industry in our country, the disease has also appeared in various provinces and cities, causing serious economic losses to the pig industry in my country and the world. The disease can break out among pigs of various ages, especially in piglets before weaning. In some areas, the incidence rate is as high as 87.97%, and the fatality rate reaches 89.90%. After infectio...

Claims

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Application Information

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IPC IPC(8): A61K31/351A61P31/14
CPCA61K31/351A61P31/14Y02A50/30
Inventor 王旭郑友乐谢胜松潘源虎陶燕飞程古月郝海红谢长清刘振利
Owner HUAZHONG AGRI UNIV
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