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Application of ethoxysanguinarine to pharmacy

A technology of ethoxysanguinarine and medicinal salts, which is applied in drug combinations, respiratory diseases, diseases, etc., to achieve significant curative effect, moderate dosage and moderate cost

Inactive Publication Date: 2012-11-21
YUNNAN AGRICULTURAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report about the anti-PRRSV infection effect and mechanism of ethoxysanguinarine in the prior art

Method used

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  • Application of ethoxysanguinarine to pharmacy
  • Application of ethoxysanguinarine to pharmacy
  • Application of ethoxysanguinarine to pharmacy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] When the Marc-145 cells on the 96-well cell plate (Corning Company) grew to a dense monolayer, the compound monomer ethoxysanguinarine EOSN (obtained by crystallization of sanguinarine in ethanol) was prepared with DMEM culture medium. 2-fold serial dilution, 5 gradients were serially diluted; 3 cell wells were inoculated in each gradient, the total amount of the mixed solution was 100 μl, and normal cell control was set at the same time; after 3 days of culture in a carbon dioxide incubator at 37°C, add 2-(2- CCK of methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid phenyl)-2H-tetrazole monosodium salt (WST-8) -8 Detection reagent, incubate for another 4 hours, then measure its absorbance at 450nm (OD450) with a microplate reader, and use the Reed-Much method to obtain the half-toxicity concentration TC of EOSN to cells according to the OD value 50 (50% toxic concentration) and the maximum non-toxic dose of TC 0 (0% toxic concentration) values ​​were 53.1...

Embodiment 2

[0030] Inoculate Macr-145 cells in a 96-well cell plate to make it cover a single layer; make 2-fold serial dilutions of EOSN, and then mix different concentrations of extracts with 300 TCID 50The virus solution is mixed, and then the virus drug mixture is added to the cells. At the same time, a normal cell control group (without drug and virus) and a virus control group (only containing virus) are set up, and the culture is continued in a 5% CO2 incubator at 37°C. Daily observation of viral CPE (cytopathic effect), the CPE of Marc-145 cells caused by PRRSV mainly caused cells to shrink, aggregate, and finally fall off; after 72 hours of culture, the difference in CPE changes in different treatment groups suggested that EOSN was in the concentration range of 7.9-31.6μM It can inhibit the proliferation of PRRS virus in Marc-145 cells, significantly reduce the damage effect (CPE) of the virus on cells, and the effect is dose-dependent ( figure 2 ); according to the CPE changes ...

Embodiment 3

[0032] The real-time RT-PCR method was used to detect the effect of drugs on the expression of PRRSV ORF7 gene mRNA, the RNA extraction reagent RNAisoTMPlus was used to extract the total RNA of the drug group and the virus control group, and the extracted total RNA was extracted using the iQ5 real time PCR instrument (Bio-Rad ) to carry out purpose amplification, according to the β-action gene and the CT value of PRRSV ORF7 gene obtained by fluorescence quantification, use the Pfaffl method to calculate the expression ratio of the ORF7 gene of the experimental group relative to the virus control group. The results showed that: when the compound EOSN concentrations were 7.9, 15.8, and 31.6 μM, the expression of ORF7 mRNA of PRRS virus was significantly lower than that of the virus control group (P image 3 shown.

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Abstract

The invention discloses application of ethoxysanguinarine to preparation of a medicament for treating virus diseases of animals, in particular to application of the ethoxysanguinarine to preparation of medicaments for treating porcine reproductive and respiratory syndrome virus infection and porcine reproductive and respiratory syndrome, preparation of a virus gene expression inhibitor as well aspreparation of medicaments of antiviral drugs and medicaments for inhibiting porcine reproductive and respiratory syndrome virus. The ethoxysanguinarine has the functions of remarkably lowering nuclear protein gene expression of the porcine reproductive and respiratory syndrome virus and inhibiting viral multiplication; and in vitro cell pharmacological experiment shows that the ethoxysanguinarine has definite anti-virus effect. The ethoxysanguinarine has wide application prospect in treatment and control of porcine reproductive and respiratory syndrome virus infection. The monomer has an easily-obtained source, low cost and higher market value.

Description

technical field [0001] The invention belongs to the technical field of medicines, and specifically relates to the application of ethoxysanguinarine in the preparation of medicines for treating viral diseases in animals, especially in the preparation and treatment of porcine reproductive and respiratory syndrome virus infection, and in the preparation of medicines for treating porcine reproductive and respiratory syndrome. Syndrome drugs, or in the preparation of viral gene expression inhibitors, in the preparation of antiviral drugs, and in the application of drugs for inhibiting PRRS virus. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS), also known as porcine blue ear disease, is a reproductive disorder in pigs caused by porcine reproductive and respiratory syndrome virus (PRRSV) and respiratory infectious diseases. Clinically, the main characteristics are reproductive disorders of sows and respiratory symptoms of piglets. Reproductive d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/4741A61P31/12A61P11/00A61P15/00
Inventor 尹革芬程永现毕峻龙吕青杨贵树李想
Owner YUNNAN AGRICULTURAL UNIVERSITY
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