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Application of shikonin to preparation of rotavirus infection resisting drugs

A technology of rotavirus and shikonin, which can be used in antiviral agents, antitumor drugs, drug combinations, etc., can solve the problems of vaccine virus shedding, vaccine cost, efficacy and safety restrictions and promotion, so as to reduce the degree of diarrhea, No toxic side effects, good safety effect

Active Publication Date: 2019-10-15
东莞市松山湖中心医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the attenuated activity of the vaccine raises some issues related to the risk of virus shedding and transmission, as well as the cost, efficacy and safety of the vaccine, which limit its promotion.

Method used

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  • Application of shikonin to preparation of rotavirus infection resisting drugs
  • Application of shikonin to preparation of rotavirus infection resisting drugs
  • Application of shikonin to preparation of rotavirus infection resisting drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Toxicity Evaluation of Shikonin on MA104 Cells

[0037] This example studies the cytotoxicity of shikonin to normal cell MA104 cell line (monkey embryonic kidney cells). The specific operation method is: take the MA104 cell line in the exponential growth phase, and make cell suspension after trypsinization and detachment After cell counting, dilute to 1×10 with high glucose DMEM containing 10% FBS 4 / mL of cell suspension, add 100 μL of cell suspension to each well of a 96-well culture plate, each well contains about 8,000 cells; suck out the medium after the cells adhere to the wall, and add shikonin liquid with different final concentrations in each experimental group (The solvent is DMEM culture fluid), respectively 0 μM (control group), 5 μM, 10 μM, 20 μM, 40 μM, 80 μM, 160 μM, 320 μM, and the control group is the same volume of DMEM culture fluid. Set 6 parallel wells for each dose in the experimental group, and set 6 parallel wells for the control wells; place at...

Embodiment 2

[0040] Inhibitory effect of shikonin on the replication and amplification of human rotavirus

[0041] This example studies whether shikonin has inhibitory effect on the replication and amplification of human rotavirus. 4 Cells / well were seeded in 96-well plates and incubated at 37°C for 24 h. After the cells adhered to the wall, the cell culture plate was taken out and washed once with PBS. The rotavirus solution with an MOI of 0.01 (100TCID50RV-Wa strain virus solution, donated by the Institute of Immunology, Third Military Medical University) was reacted with 10 μg / mL trypsin for 30 minutes, and then added to a 96-well culture plate, 100 μl / well. The normal cell control group was added with an equal volume of DMEM medium. All cells at 37°C, 5% CO 2 After incubation for 2 hours, the virus liquid was aspirated, and shikonin liquid of different concentrations (0 μM (virus control group), 5 μM, 10 μM, 20 μM, 40 μM, 80 μM) was added, 100 μL / well. Add an equal volume of DMEM c...

Embodiment 3

[0044] Inhibitory Effect of Shikonin on Antigen Synthesis of Human Rotavirus

[0045] This example studies whether shikonin has inhibitory effect on the synthesis of human rotavirus antigen. 5 Cell density seeded in 6-well plates at 37°C, 5% CO 2 cultivated under conditions. After the cells were completely adhered to the wall, the old culture medium in the cells was removed, and serum-free DMEM culture medium was added to the 6-well plate for 6 h. After 6 hours, the DMEM medium without FBS was aspirated, and the cells were added to 100 TCID50RV-Wa strain virus solution incubated with 10 μg / ml trypsin, and cultured at 37°C for 2 hours. After 2 hours, suck out the virus liquid, wash the cells with DMEM culture medium without FBS, add shikonin liquid at different concentrations (5 μM, 10 μM, 20 μM, 40 μM, 80 μM), and add an equal volume of DMEM without FBS to the control group Medium, continue to culture at 37°C for 24h. After 24 hours, collect the cell culture medium separat...

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Abstract

The invention relates to application of shikonin to preparation of rotavirus infection resisting drugs. The shikonin, or pharmacologically acceptable salt, ester and solvate thereof finally achieve the beneficial effect of rotavirus resistance by inhibiting replication and amplification of rotaviruses and inhibiting antigen synthesis of the rotaviruses; and the shikonin, or the pharmacologically acceptable salt, ester and solvate thereof have no toxic and side effect on normal cells, are good in safety, provide a theoretical basis for studying the treatment strategy of rotavirus infection, andprovide an embedding point for preparation of the new rotavirus-resisting drugs.

Description

technical field [0001] The invention relates to the field of medicine application, in particular to the application of shikonin in the preparation of medicines against rotavirus infection. Background technique [0002] Rotavirus (RV), a member of the Reoviridae family, is the main pathogen that causes viral diarrhea. About 110 million newborns around the world suffer from severe secretory diarrhea due to RV infection every year, and about 600,000 children die of dehydration due to RV infection. Compared with other viral or bacterial infectious diarrhea, the degree and duration of vomiting and diarrhea caused by rotavirus are much longer than other types of diarrhea, which has a huge impact on human health and economy. Vaccination remains the most effective means of preventing RV infection. [0003] CN106676076A discloses a method for preparing a stock solution of rotavirus vaccine from serum-free Vero cells, comprising the following steps: cultivating Vero cells with serum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/122A61K31/222A61P31/14A61P35/02A61P1/12
CPCA61K31/122A61K31/222A61P31/14A61P35/02A61P1/12
Inventor 黄浩海蒲荣廖丹
Owner 东莞市松山湖中心医院
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