Application of biapenem in preparation of medicine for preventing and treating bovine enterovirus infection

A technology of biapenem and enterovirus, applied in the field of application of biapenem in the preparation of drugs for preventing and treating bovine enterovirus infection

Active Publication Date: 2020-02-21
DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Application of biapenem in preparation of medicine for preventing and treating bovine enterovirus infection
  • Application of biapenem in preparation of medicine for preventing and treating bovine enterovirus infection
  • Application of biapenem in preparation of medicine for preventing and treating bovine enterovirus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Virus TCID 50 Determination of

[0051] MDBK cells (preserved by Dairy Cow Research Center, Shandong Academy of Agricultural Sciences) were digested and divided into 3 × 10 cells per well. 5 Cells / mL were seeded into 96-well cell culture plates, placed in 37°C, 5% CO 2 After being cultivated into monolayer cells in the cell incubator, discard the cell growth solution in the well, and the virus dilution solution of bovine enterovirus serial 10-fold dilution (dilution degree is 10 -1 ~10 -10 ) inoculated in a 96-well plate full of monolayer cells, 100 μL per well, placed in 37°C, 5% CO 2 Continue to culture in the incubator, observe the CPE of the cells day by day, and record the number of cells with pathological changes in detail. At the same time, a normal cell control group and a blank control group were set up, with 8 replicates in each group, and the results were judged when no further cell lesions occurred. Cytopathic wells are cell wells correspondin...

Embodiment 2

[0062] Example 2 Toxicity test of biapenem on MDBK cells:

[0063] MDBK cells are susceptible cells to bovine enterovirus. Therefore, the cytotoxicity of biapenem to MDBK cells was first detected, and the specific experimental steps were as follows:

[0064] (1) Inoculate 100 μL cells (MDBK 3×10 4 pieces / hole).

[0065] (2) After culturing for about 12 hours, the next step of drug addition analysis was carried out. Discard the medium, add 100 μL of 2% FBS DMEM containing different drug concentrations to each well, and make 3 parallels for each concentration. At the same time, control wells: add 100 μL 2% FBS DMEM medium. Zero well: no cells are plated.

[0066] (3) At 37°C, 5% CO 2 After culturing under the conditions for 48 hours, operate according to the instructions of the CCK-8 kit, and measure the OD value at 450nm with a microplate reader.

[0067] (4) 37°C, 5% CO 2 After culturing for 4 h under the same conditions, the absorbance was measured at 450 nm. A450nm ...

Embodiment 3

[0070] Example 3 Inhibition experiment of biapenem on bovine enterovirus:

[0071] (1) Inoculate 3×10 in each well of a 96-well plate 4 MDBK cells, 37°C, 5% CO 2 Cultivate overnight in an incubator;

[0072] (2) Discard the medium and add 100 μL 100TCID to each well 50 The bovine enterovirus dilution solution (use 2% FBS DMEM to add the virus dilution solution after the cells are overgrown, according to the initial concentration of 100 μ M, double the concentration gradient dilution and dosing, 5% CO 2 Cultivated in an incubator;

[0073] (3) After 48 hours, operate according to the instructions of the CCK-8 kit, and measure the OD value at 490 nm with a microplate reader.

[0074] (4) analysis data, virus inhibition rate (%)=(drug treatment group D450nm value-virus control group D450nm value) / (normal cell control group D450nm value-virus control group D450nm value) * 100%, get with GraphPad Prism5 software The half effective concentration of the compound (EC 50 )value. ...

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Abstract

The invention provides an application of biapenem in preparation of a medicine for preventing and treating bovine enterovirus infection. The compound biapenem can effectively inhibit proliferation ofbovine enterovirus and has low toxicity to cells, and experiments prove that the median toxicity concentration (CC50) of biapenem to MDBK cells is greater than 100 [mu]M, and the median effective concentration (EC50) of biapenem to bovine enterovirus is 60 [mu]M; the treatment index of biapenem to bovine enterovirus is greater than 1.67, which indicates that biapenem has a prospect of being developed into a drug for preventing and/or treating bovine enterovirus infection, opens up the new drug application for biapenem, lays an experimental foundation for developing efficient and specific anti-BEV drugs, and provides a new visual field.

Description

technical field [0001] The invention belongs to the technical field of medicines and drugs, and in particular relates to the application of biapenem in the preparation of drugs for preventing and treating bovine enterovirus infection. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Bovine Enterovirus (BEV) is a member of the family Picornaviridae and the genus Enterovirus. Picornavirus is an extremely complicated virus family, which can be divided into 5 genera including rhinovirus, foot-and-mouth disease virus, enterovirus, cardiovirus and hepatitis A virus. There are more than 200 kinds of viruses in total, which can cause human and animal Many diseases, such as hepati...

Claims

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Application Information

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IPC IPC(8): A61K31/4196A61P31/14
CPCA61K31/4196A61P31/14Y02A50/30
Inventor 程凯慧楚会萌杨宏军任亚初解晓莉张亮于志君孙阳阳朱彤
Owner DAIRY CATTLE RES CENT SHANDONG ACADEMY OF AGRI SCI
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