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Application of reserpine in preparing anti-porcine epidemic diarrhea virus medicine

A technology for porcine epidemic diarrhea and reserpine, which is applied in antiviral agents, pharmaceutical formulations, organic active ingredients, etc., can solve the problem that virus particles do not have hemagglutination, and achieve the effects of easy absorption, low drug, and significant effect.

Active Publication Date: 2018-09-14
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, PEDV is sensitive to fat-soluble fluids, and the virion does not have hemagglutination

Method used

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  • Application of reserpine in preparing anti-porcine epidemic diarrhea virus medicine
  • Application of reserpine in preparing anti-porcine epidemic diarrhea virus medicine
  • Application of reserpine in preparing anti-porcine epidemic diarrhea virus medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Reserpine Cytotoxicity Determination on Vero-E6 Cells

[0028] After counting Vero-E6 cells, they were diluted with 8% fetal bovine serum (FBS) DMEM nutrient solution to an appropriate density and then diluted to 1.5×10 4 The concentration per well was added to a 96-well plate and placed at 37°C, 5% CO 2 After the cells adhered to form a monolayer in the incubator (about 18-20h), the reserpine was diluted with DMEM nutrient solution to: 1 μM, 2 μM, 5 μM, 10 μM, and three sets of repetitions were set for each concentration. After treating Vero-E6 cells with 100 μl / well of DMSO control and diluted reserpine samples on a 96-well cell culture plate for 72 hours, remove the supernatant, add 90 μl of fresh culture medium, and then add 10 μl of MTT solution, and continue to incubate for 4 hours; Remove the supernatant, add 110 μl Formazan solution to each well, place on a shaker and shake at low speed for 10 minutes to fully dissolve the crystals. Measure the abs...

Embodiment 2

[0030] Example 2 Western Blot Determination of Reserpine Inhibiting Porcine Epidemic Diarrhea Virus Infection on Vero-E6 Cells Activity

[0031] After Vero-E6 cells were counted, they were diluted with 8% fetal bovine serum (FBS) DMEM nutrient solution to an appropriate density and then diluted to 7×10 5 The concentration per well was added to a 6-well plate and placed at 37°C, 5% CO 2 After the cells adhered to form a monolayer in the incubator and grew to a density of 70-80% (about 18-20 hours), the reserpine was diluted with DMEM nutrient solution to: 1 μM, 2 μM, 5 μM, 10 μM. DMSO control and different concentrations of reserpine were pre-treated Vero-E6 cells at 37°C for 2 hours, then infected with porcine epidemic diarrhea virus HLJBY strain (MOI=0.01), and placed at 37°C, 5% CO 2 Place in the incubator for 1 hour, wash three times with PBS, add 1ml of DMEM maintenance solution containing 2% fetal bovine serum (FBS) and contain reserpine, place at 37°C, 5% CO 2 Afte...

Embodiment 3

[0032] Example 3 Fluorescent quantitative detection of the activity of reserpine in inhibiting porcine epidemic diarrhea virus infection on Vero-E6 cells sex

[0033] After Vero-E6 cells were counted, they were diluted with 8% fetal bovine serum (FBS) DMEM nutrient solution to an appropriate density and then diluted to 7×10 5 The concentration per well was added to a 6-well plate and placed at 37°C, 5% CO 2After the cells adhered to form a monolayer in the incubator and grew to a density of 70-80% (about 18-20 hours), the reserpine was diluted to 10 μM with DMEM nutrient solution. After pre-treating Vero-E6 cells with DMSO control and reserpine at 37°C for 2 hours, they were infected with porcine epidemic diarrhea virus HLJBY strain (MOI=0.01), and placed at 37°C, 5% CO 2 Place in the incubator for 1 hour, wash three times with PBS, add 1ml of DMEM maintenance solution containing 2% fetal bovine serum (FBS) and contain reserpine, place at 37°C, 5% CO 2 After culturing in...

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Abstract

The invention discloses application of reserpine in preparing anti-porcine epidemic diarrhea virus medicine. The mass percent of the reserpine is not less than 99.40%, and the application dose is 1-10micromoles per liter; the effect of the reserpine on the porcine epidemic diarrhea virus is very obvious, the medicine is safe and has few toxic and side effects, the drug residue is low, and no pollution exists.

Description

technical field [0001] The invention belongs to the field of antiviral applications, and in particular relates to the application of reserpine in the preparation of anti-porcine epidemic diarrhea virus drugs. Background technique [0002] Porcine epidemic diarrhea is a disease caused by porcine epidemic diarrhea virus (porcine epidemic diarrhea virus, PEDV). It is clinically characterized by vomiting, severe diarrhea and dehydration. Infect. Among them, suckling piglets are the most susceptible, the incidence rate can reach 100%, and the case fatality rate reaches 80%-100%. Back-up pigs, sows or fattening pigs are transient diarrhea, and the incidence rate is lower. PEDV first appeared in Europe in the 1970s. However, since 2010, the outbreak of PEDV mutant strains has caused huge economic losses to the pig industry in China, Japan, South Korea, Thailand, the Philippines and other Asian countries. Pandemic in the United States, Germany, Portugal and other countries, the pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/475A61P31/14
CPCA61K31/475A61P31/14
Inventor 钱莺娟郑龙三陈新薄宗义
Owner NANJING AGRICULTURAL UNIVERSITY
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