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Application of artemisinin and derivative thereof in preparation of drug for preventing and treating porcine reproductive and respiratory syndrome

A technology for respiratory syndrome and derivatives, which is applied in the direction of pharmaceutical formulas, medical preparations containing active ingredients, organic active ingredients, etc., can solve the problems of unsatisfactory vaccine prevention and control effects, inability to stimulate cellular immunity, and long-term effects, and achieve drug Effective components and mechanism of action are clear, and the preparation process is mature and reliable.

Inactive Publication Date: 2018-07-24
SOUTH CHINA AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 1996, the PRRS pathogen was isolated from aborted pigs in my country; in 2006, a highly pathogenic PRRS disease characterized by high mortality of piglets broke out in China, which caused huge losses to my country's pig industry and was listed as a first-class disease in my country. Animal diseases, and the effect of vaccine control is not ideal
At present, the situation of prevention and control of highly pathogenic PRRS is very serious. The virus is prone to recombination, immune escape and other problems, and it is easy to trigger secondary infections such as swine fever virus, porcine circovirus, and swine influenza virus. There is no effective control method yet, and vaccination is the main preventive measure
However, the main problems in the clinical use of PRRS vaccines are: first, it takes a long time for the vaccine to produce an immune response; second, the PRRSV vaccine does not have complete protection, especially the inactivated vaccine cannot stimulate cellular immunity; third, PRRSV The attenuated vaccine can persist in the tissues of immunized pigs for several weeks and spread from vaccinated pigs to susceptible pigs
At present, there is no report on the anti-porcine reproductive and respiratory syndrome virus effect of artemisinin and its derivatives, and there are no published related patents

Method used

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  • Application of artemisinin and derivative thereof in preparation of drug for preventing and treating porcine reproductive and respiratory syndrome
  • Application of artemisinin and derivative thereof in preparation of drug for preventing and treating porcine reproductive and respiratory syndrome
  • Application of artemisinin and derivative thereof in preparation of drug for preventing and treating porcine reproductive and respiratory syndrome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Effects of artemisinin and its derivatives on the viability of Marc-145 cells and PAM cells

[0025] After inoculating Marc-145 cells or PAM cells in a 96-well cell culture plate and growing to a monolayer, the medium was discarded, washed twice with PBS (phosphate buffered saline), and the drug was treated with a cell maintenance solution containing 2% fetal bovine serum. Dilute to the concentration to be tested, add 100 μL / well, and make three replicate wells for each concentration. After continuing to incubate at 37°C for 48h, add 20μL of 5mg / ml MTT reagent to each well, incubate at 37°C for 4h, discard the supernatant, add 150mL DMSO, shake for 10min to fully dissolve the formed formazan crystals, and detect at 490nm wavelength absorbance value. from figure 1 It can be seen that at the concentration of 2.5 μg / mL of artemisinin and its derivatives, the survival rate of Marc-145 cells and PAM cells is more than 80%, and the growth of cells is not affected...

Embodiment 2

[0026] Example 2: Indirect immunofluorescence assay (IFA) analysis of the inhibition of PRRSV virus N protein synthesis after treatment of Marc-145 cells with different concentrations of artemisinin and its derivatives

[0027] After the Marc-145 cells were seeded in a 6-well cell culture plate and grown to a monolayer, the medium was discarded, washed twice with PBS (phosphate buffered saline), and diluted with DMEM cell maintenance solution containing 2% fetal bovine serum. with 100 TCID 50 of porcine reproductive and respiratory syndrome virus (PRRSV), 1 mL / well, incubated at 37°C for 2 h, discarded the virus supernatant, washed twice with PBS, and added 2.5, 1.25, and 0.6 μg / mL of drugs, 2 mL / well. In the experiment, the normal control group (without the test drug and PRRSV) and the PRRSV control group (without the test drug) were set at the same time, and three parallels were set for each test concentration. The culture was terminated after continuing to culture at 37°C ...

Embodiment 3

[0028] Example 3: Western Blot analysis of changes in PRRSV virus N protein expression after treatment of Marc-145 cells with different concentrations of artemisinin and its derivatives

[0029] The steps of infecting Marc-145 cells with PRRSV and adding drugs are the same as in step 2. After the cells are cultured at 37° C. for 48 h, the supernatant is discarded and washed twice with PBS. Place the cell culture plate on ice, add 150 μL / well of RIPA lysis solution, and after lysing by pipetting, aspirate the liquid into a centrifuge tube, centrifuge at 12,000 rpm for 5 minutes, and aspirate the supernatant into another clean tube for use. The protein concentration of each sample was determined according to the method of the BCA protein concentration assay kit. The bands of N protein of PRRSV and internal reference protein GAPDH were detected by Western Blot. The test results are as image 3 The results showed that artemisinin and its derivatives had different degrees of inhi...

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Abstract

The invention discloses application of artemisinin and a derivative thereof in preparation of a drug for preventing and treating the porcine reproductive and respiratory syndrome. A pharmacological test shows that the artemisinin and the derivative thereof have an outstanding protection effect on cells infected with a porcine reproductive and respiratory syndrome virus, and has obvious activity ofresisting the porcine reproductive and respiratory syndrome virus. The artemisinin and the derivative thereof can be used as effective components to prepare a new drug for preventing and treating theporcine reproductive and respiratory syndrome, and have a wide application prospect.

Description

technical field [0001] The invention relates to the field of veterinary medicine, in particular to the preparation of a medicine for preventing and treating porcine reproductive and respiratory syndrome. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS), commonly known as "porcine blue-ear disease", is a porcine reproductive and respiratory syndrome virus (PRRSV) caused by porcine reproductive and respiratory syndrome virus (PRRSV). The new catastrophic disease is a highly contagious disease with clinical manifestations of severe reproductive failure in sows, widespread pneumonia, growth retardation and increased mortality in weaned pigs. PRRSV is a single-stranded RNA virus of the Arteriviridae family, belonging to the order Mantovirus, prone to genetic recombination and low cross-protection of strains. Its strains are divided into European strains and American strains. The widespread PRRS in my country is caused by the variation of America...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/366A61K31/357A61P31/14
CPCA61K31/357A61K31/366
Inventor 陈建新田鸽李志萍刘雅红邓衔柏邱电吴倩倩张明昕
Owner SOUTH CHINA AGRI UNIV
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