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Application of toosendanin in preparation of medicine for preventing and treating porcine virus infectious diseases

A technology for infectious diseases, toosendanin, is applied in the field of use in the preparation of drugs for preventing and treating swine virus infectious diseases, and can solve the problems of frequent occurrence of mutant strains, vaccine lag, economic losses, etc., and is suitable for large-scale industrialization. The effect of production use, wide source and high safety

Pending Publication Date: 2022-06-07
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because PEDV infection can lead to high mortality of suckling piglets and weaned piglets, the mortality rate of piglets at one week old is almost 100%, thus causing huge economic losses to the pig industry in many countries
Vaccines are one of the important means of controlling pandemic diseases. However, because PEDV is prone to mutations under the pressure of field selection, new mutant strains frequently appear, which makes vaccines have serious lags. Therefore, it is urgent to develop effective broad-spectrum anti-PEDV drugs
[0007] In recent years, there have been more and more research reports on anti-PRRSV, ASFV and PEDV drugs, but most of the related research stays at a relatively primary level, and the effective concentration of active compounds is high, and the mechanism of action has not been elucidated clearly, so it is difficult to achieve the desired effect as a drug. The four basic requirements of "safety, effectiveness, stability, and controllability" are necessary, and the druggability is poor

Method used

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  • Application of toosendanin in preparation of medicine for preventing and treating porcine virus infectious diseases
  • Application of toosendanin in preparation of medicine for preventing and treating porcine virus infectious diseases
  • Application of toosendanin in preparation of medicine for preventing and treating porcine virus infectious diseases

Examples

Experimental program
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preparation example Construction

[0040] Wherein, toosendanin used in the present invention is prepared by the following method:

[0041] The root bark of the neem tree was pulverized with a pulverizer and then sieved (20 mesh), and extracted twice with a 90% volume fraction of ethanol solution at 50 °C in a water bath for 2 hours each time. Neem crude extract; Weigh a certain amount of Chuan Neem crude extract, extract with petroleum ether, ethyl acetate and n-butanol in turn, distill under reduced pressure for the ethyl acetate extraction part to obtain a concentrated paste and pass through silica gel column chromatography, Elute with 1% methanol-chloroform, 2% methanol-chloroform and 5% methanol-chloroform in sequence, collect the eluent containing thuanimin, concentrate under reduced pressure, pass through a silica gel column again, and elute with 1% methanol-chloroform , the eluent was passed through Sephadex LH-20 column, and eluted with 50% methanol-water to obtain purified toosendanin, and the purity w...

Embodiment 1

[0043] Example 1 Inhibitory effect of different concentrations of toosendanin on the proliferation of ASFV DNA in PAMs cells

[0044] 1. Experimental method

[0045] Resuscitate porcine alveolar macrophages (Porcine alveolar macrophages, PAMs) (separated from pig lungs and stored in liquid nitrogen tanks in advance); use RPMI-1640 culture medium (containing 10% fetal bovine serum, 100U / mL) penicillin, 100U / mL streptomycin) to resuspend the cells, and press 8 × 10 5 Cells / well were inoculated into 24-well plates; set the Sichuansendan high (3 μM), medium (1 μM), low concentration groups (0.3 μM), virus group (Mock), and cell control group (NC, no infection, no drug administration) , 3 replicates per group; 37°C, 5% CO 2 The cells were cultured in a cell incubator for 6 hours; the monolayer of PAMs cells in a 24-well plate with a filling degree of about 80-90% was infected with 2MOI of ASFV. Serum RPMI-1640 culture medium) diluted and prepared 3 μM, 1 μM, 0.3 μM concentration...

Embodiment 2

[0052] Example 2 Inhibitory effect of different concentrations of toosendanin on ASFV RNA transcription in PAMs cells

[0053] 1. Experimental method:

[0054] To revive PAMs, resuspend cells in RPMI-1640 medium and press 8 x 10 5 Cells / well were inoculated into 24-well plates; set the Sichuanese high (3μM), medium (1μM), low concentration groups (0.3μM), virus group (Mock), and cell control group (NC), with 3 replicates in each group; 37℃, 5%CO 2 The cells were cultured in a cell incubator for 6 hours; the monolayer of PAMs cells in a 24-well plate with a filling degree of about 80-90% was infected with 2MOI of ASFV. Serum RPMI-1640 culture medium) diluted and prepared 3 μM, 1 μM, 0.3 μM concentration of toosendanin, placed at 37 ° C, 5% CO 2 The incubation was terminated after 24 hours of incubation in the incubator.

[0055] After observing the cell morphology, freeze and thaw the cell plate at -80°C and 4°C for three times to fully lyse the cells, so that all the virus...

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Abstract

The invention belongs to the technical field of veterinary drugs, and particularly relates to application of toosendanin to preparation of drugs for preventing and treating porcine virus infectious diseases. The invention provides novel antiviral application of toosendanin, which is proved by research of various methods that toosendanin can remarkably inhibit virus proliferation at a micromolar concentration level, particularly has a remarkable inhibiting effect on African swine fever virus, porcine reproductive and respiratory syndrome virus and porcine epidemic diarrhea virus, and can be used for preparing antiviral drugs. The effect of simultaneously preventing and treating African swine fever, porcine reproductive and respiratory syndrome and porcine epidemic diarrhea can be achieved. The toosendanin is wide in source, low in cost, high in clinical application safety and very suitable for large-scale industrial production and use.

Description

technical field [0001] The invention belongs to the technical field of veterinary medicines. More specifically, it relates to the use of toosendanin in the preparation of medicines for the prevention and treatment of swine virus infectious diseases. Background technique [0002] African swine fever (ASF), caused by African swine fever virus (ASFV), is a highly contagious animal disease with a high incidence. Affected pigs often show acute, febrile, and hemorrhagic clinical symptoms, and their organs show severe vascular lesions, such as lymph node hemorrhage, renal hemorrhage, disseminated intravascular coagulation, thrombocytopenia, etc. Virulent strain infection can cause 100% of the disease. Mortality rate (Kang Ya Nan et al. (2021), Current situation and prevention and control measures of African swine fever in my country, China Swine Industry, 16(4):57-60). ASF is a statutory notifiable disease of the World Organization for Animal Health (Office International Des Epizo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/58A61K36/58A61P31/12A61P31/14A61P31/20
CPCA61K31/58A61K36/58A61P31/14C07J71/001
Inventor 陈建新刘远佳张明昕亓文宝张玲华武力宋高鹏何应敏朱君海林琪胜
Owner SOUTH CHINA AGRI UNIV
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