Application of Tripterygium in the Preparation of Drugs for Inhibiting the Replication of Marek's Disease Virus

A technology for Marek's disease and tripterygne, applied in antiviral agents, pharmaceutical formulations, plant raw materials, etc., to achieve enhanced effects, broad application prospects, good economic value and research significance

Active Publication Date: 2022-04-29
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that tripterygne has various pharmacological activities such as anti-tumor, anti-inflammation, anti-oxidation, and pain relief, but so far there has been no report about tripterygne’s anti-Marek’s disease or any inhibition of MDV virus replication and activity

Method used

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  • Application of Tripterygium in the Preparation of Drugs for Inhibiting the Replication of Marek's Disease Virus
  • Application of Tripterygium in the Preparation of Drugs for Inhibiting the Replication of Marek's Disease Virus
  • Application of Tripterygium in the Preparation of Drugs for Inhibiting the Replication of Marek's Disease Virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Tripterine inhibits the enzymatic activity of UL36 encoded by MDV.

[0075] Enzyme reaction system: 1.5nM UL36 enzyme in 100μL reaction system, 1× reaction buffer (50mM HEPESpH7.8, 0.1mg / mL BSA, 0.5mM EDTA, 1mM DTT), add reaction fluorescent substrate Ub-AMC,37 Incubate at °C, and measure the change in fluorescence intensity on a fluorescence spectrophotometer. The reaction system with 2 μM tripterine was added as "inhibitor group", and the reaction system without tripterine was added with the same amount of solvent DMSO as "no inhibitor group". The result is as figure 1 , UL36 in the "no inhibitor group" hydrolyzed Ub-AMC normally and produced a fluorescent signal, while the activity of UL36 in the "inhibitor group" was inhibited, and the fluorescent signal did not increase. This result shows that tripterine inhibits the enzymatic activity of UL36 encoded by MDV.

Embodiment 2

[0077] Preparation of CEF cells and infection with MDV:

[0078] 1. Take SPF chicken embryos aged 9-11 days, wipe the chicken embryos with alcohol cotton, and wipe and disinfect from the air chamber to the surrounding. Then put the chicken embryo into the ultra-clean workbench with the air cell facing up. Carefully tap the air cell with autoclaved forceps and clip off the eggshell.

[0079] 2. Use new sterile tweezers to uncover the allantoic chorion, and discard the egg shell and vitelline membrane. Hold the head of the embryo with sterilized forceps and gently transfer it to the plate.

[0080] 3. Fix the head with sterilized tweezers, cut off the head, limbs, and organs with sterilized scissors, and rinse repeatedly with sterilized PBS until colorless.

[0081] 4. Use sterilized scissors to cut the tissue pieces until there are no large particles of tissue, add 8 mL of sterilized PBS to wash, let stand for 1 min, and discard the suspension mixed with red blood cells and ...

Embodiment 3

[0098] Tripteryglide inhibits the replication of MDV and the occurrence of MD at the level of chicken MD disease model:

[0099] The specific operation is: take 1-day-old SPF chickens, according to the MDV titer of 1000PFU, the way of attacking the virus is intraperitoneal inoculation, and these challenged chickens are treated with intraperitoneal injection of tripterine every day, and the experiment is divided into 0.01, 0.05, 0.1, 0.15 , 0.2, 0.3 and 0.5, a total of 7 groups, no drug treatment, only injected with the same amount of DMSO as the control group. 10 chickens per group. 20 days after the challenge, blood was collected from the blood vessels under the wings, T lymphocytes were separated, and the quantitative analysis of the MDV gene copy number was carried out according to the number of 200,000 cells, as shown in Figure 7 . The MD incidence rate of each group chicken in 50 days after the statistical challenge, such as Figure 8 . 50 days after the challenge or...

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Abstract

The invention provides a medicine for inhibiting the replication of Marek's disease virus, including a deubiquitinase UL36 inhibitor, the deubiquitinase UL36 inhibitor being tripterygium twig extract, and a medicine for preventing or resisting Marek's disease medicine. Experiments of inhibiting virus-encoded enzyme activity, drug toxicity experiments at the CEF cell level, virus CPE inhibition experiments and chicken MD model experiments proved that the tripterygium wilfordii extract can effectively inhibit the replication of Marek's disease virus and the growth of chicken MD tumors. Produced, it has a good application prospect in the poultry breeding industry.

Description

technical field [0001] The invention relates to the application field of natural compounds, in particular to the application of Tripterygium wilfordii extract as a drug for inhibiting the replication of Marek's disease virus and preventing the occurrence of Marek's disease. Background technique [0002] Marek's disease (MD) is a highly contagious, highly pathogenic, lymphoproliferative viral neoplastic disease caused by chickens infected with the pathogenic serotype I type Marek's disease virus (MDV), which is spread all over the world Widely exist, my country and Egypt are countries with high incidence of this disease. Therefore, MD is an important threat to my country's poultry industry. Chicken is the most important natural host of MDV, which belongs to the cell-associated herpesviridae family, and its molecular structure and gene composition are similar to alphaherpesviruses. Unless kept under strict Specific Pathogen Free (SPF) conditions, all flocks can become infecte...

Claims

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Application Information

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IPC IPC(8): A61K31/56A61K36/37A61K31/585A61K31/436A61P31/22
CPCA61K36/37A61K31/56A61K31/585A61K31/436A61P31/22
Inventor 艾永兴许家翠李占军徐媛媛
Owner JILIN UNIV
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