A drug that inhibits the replication of Marek's disease virus, prevents or fights Marek's disease

A technology for Marek's disease and virus replication, applied in antitumor drugs, antiviral agents, drug combinations, etc., can solve the problems of no MDV virus replication, no reports on pharmacological functions, etc., and achieve good economic value and research significance, Enhanced effect, strong effect

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

Its pharmacological function has not been reported so far, and so far there has not been any report about DUBs-IN-3 anti-MD or any inhibition of MDV virus replication

Method used

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  • A drug that inhibits the replication of Marek's disease virus, prevents or fights Marek's disease
  • A drug that inhibits the replication of Marek's disease virus, prevents or fights Marek's disease
  • A drug that inhibits the replication of Marek's disease virus, prevents or fights Marek's disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] DUBs-IN-1 inhibits the enzymatic activity of UL36 encoded by MDV.

[0044] Enzyme reaction system: 2.5nM UL36 protein 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 added with 5 μM DUBs-IN-1 was used as "inhibitor group", and the reaction system without DUBs-IN-1 but added with the same amount of solvent DMSO was used 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 indicated that DUBs-IN-1 inhibited the enzymatic activity of UL36 encoded by MDV.

Embodiment 2

[0046] DUBs-IN-1 inhibits MDV replication at the level of CEF cells.

[0047] Preparation of CEF cells and infection with MDV:

[0048] 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.

[0049] 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.

[0050] 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.

[0051] 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 sta...

Embodiment 3

[0068] DUBs-IN-1 inhibits the replication of MDV and the occurrence of MD at the level of chicken MD disease model.

[0069] The specific operation is: take 1-day-old SPF chickens, according to the MDV titer of 1000PFU, the way of challenge is intraperitoneal inoculation, these challenge chickens are treated with intraperitoneal injection of DUBs-IN-1 every day, and the experiment is divided into 0.001, 0.005, 0.01, 0.04, 0.1, 0.15 and 0.2, 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 when they were o...

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Abstract

The invention provides a drug for inhibiting the replication of Marek's disease virus, including deubiquitinase UL36 inhibitor, and a preventive or anti-Marek's disease drug. Through the experiment of inhibiting the enzyme activity encoded by the virus, the drug toxicity experiment at the CEF cell level, the virus CPE inhibition experiment and the chicken Marek's disease model experiment, the present invention screens out the ubiquitinase UL36 inhibitor DUBs‑IN‑1 and its derivatives for the first time It can not only inhibit the activity of UL36 at the enzyme level, inhibit the replication of Marek’s disease virus at the cell level, but also inhibit the growth of Marek’s disease tumors and visceral hemorrhage at the animal level, and has a strong effect and a small dose Advantages, it can be widely used as an anti-MD drug in the production practice of poultry industry to prevent Marek's disease.

Description

technical field [0001] The invention relates to the application field of compounds, in particular to the application of DUBs-IN-1 and its derivatives as drugs for inhibiting the replication of Marek's disease virus and for 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 infected, eve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/498A61P31/22A61P35/00
CPCA61K31/498A61P31/22A61P35/00A61K2300/00
Inventor 艾永兴李占军许家翠徐媛媛
Owner JILIN UNIV
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