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Application of sophoridine in preparation of anti-herpes virus medicine

A technology of herpes virus and herpes simplex virus, which is applied in the direction of antiviral agents, drug combinations, diseases, etc., and can solve problems such as complex components, unstable active ingredient content and activity intensity

Pending Publication Date: 2021-08-24
CHENGDU UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the medicine for treating oral herpes simplex disclosed in this application, due to the complex composition of the active ingredient Sophora sophora, the problem of unstable active ingredient content and activity intensity is prone to occur.

Method used

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  • Application of sophoridine in preparation of anti-herpes virus medicine
  • Application of sophoridine in preparation of anti-herpes virus medicine
  • Application of sophoridine in preparation of anti-herpes virus medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Plaque test to detect the inhibitory effect of drugs on viral plaques

[0038] 1. Experimental method

[0039] Determination of virus titer: Vero cells in the logarithmic growth phase were taken, digested with trypsin and centrifuged, counted, and counted according to 1.3×10 5 Seed / ml density to 96-well plate, 100 μl per well, place at 37°C, 5% CO 2 cultured in an incubator. Discard the supernatant after the cells grow into a monolayer, and dilute the virus stock solution to 10 times with the maintenance solution. -1 to 10 -9 For a series of gradients, 6 replicate wells were set for each concentration, and 100 μl of virus dilution per well was inoculated on Vero cells, at 37°C, 5% CO 2 Adsorb in the incubator for 2 hours, discard the supernatant, add 100 μl of maintenance solution to each well to continue the culture, and set a normal cell control group. The cytopathic effect (CPE) was observed and recorded under an inverted microscope every day, and the...

Embodiment 2

[0043] Example 2: Plaque test method to detect the influence of drugs on the virulence of progeny viruses

[0044] 1. Experimental method

[0045] Seed Vero cells on a 12-well plate at an appropriate density. After the cells are fused the next day, add HSV-1 to infect the cells. Place them in a 5% CO2 culture at 37°C for 2 hours. Discard the supernatant and add different concentrations of The cells were treated with sophoridine and placed in the incubator for 24 hours. After culturing for 24 hours, discard the supernatant and add 1ml of maintenance solution to each well, transfer the 12-well plate to -80°C and 37°C for three times of freezing and thawing to release virus particles, collect the supernatant and place in Standby at -80°C. The virus titer of progeny was determined by plaque test.

[0046] 2. Experimental results

[0047] Plaque test results such as image 3 As shown, it can be seen that sophoridine has a significant inhibitory effect on the generation of HSV-...

Embodiment 3

[0048] Embodiment 3: Western blotting test detects the impact of drugs on viral protein gB, gD expression

[0049] 1. Experimental method

[0050] Inoculate vero cells or Hela cells in 6-well plates, grow into a single layer, and use 100TCID 50 Infect the cells with HSV-1 virus liquid, absorb in a 37°C, 5% CO2 incubator for 2 hours, discard the supernatant, add gradient diluted sophoridine solutions of different concentrations after washing with PBS, and culture in a 37°C, 5% CO2 incubator 24h. After 24 hours, the supernatant was discarded, and pre-cooled PBS was added to wash twice, each time for 3 minutes. Add 120 μL of RIPA lysate (containing 1% protease inhibitor and 1% phosphatase inhibitor), and lyse on ice for 30 min. During this period, gently pipette with a pipette to make the lysate fully contact with the cells. After fully lysed, centrifuge at 12000g, 4°C for 15min. The supernatant was taken, and the total protein content was determined using the BCA protein qua...

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Abstract

The invention provides application of sophoridine in preparation of anti-herpes virus drugs, and belongs to the field of pharmacy. Experimental results show that sophoridine can effectively inhibit formation of HSV-1 virus plaques and significantly inhibit generation of filial generation viruses; the sophoridine can also significantly inhibit the expression levels of gB and gD proteins of the HSV-1 virus, and significantly inhibit the expression levels of UL27 and US6 genes. The sophoridine has a remarkable inhibiting effect on the growth of the herpes simplex virus type 1, can be used for preparing the medicine for resisting the herpes simplex virus type 1, and can also be used for preparing the medicine for treating the infectious diseases of the herpes simplex virus type 1. In addition, sophoridine has low toxicity to normal cells, and has wide application prospects in preparation of anti-herpes virus drugs and drugs for prevention and treatment of herpes virus infection diseases.

Description

technical field [0001] The invention belongs to the field of pharmacy, and in particular relates to the application of sophoridine in the preparation of anti-herpes virus medicines. Background technique [0002] Herpes simplex virus (HSV) belongs to the family Herpesviridae and is a typical representative of herpes virus. Herpes simplex virus can cause a variety of diseases in humans, such as gingivostomatitis, keratoconjunctivitis, encephalitis, reproductive system infection and neonatal infection. According to the antigenicity of herpes simplex virus, it is divided into type 1 (HSV-1) and type 2 (HSV-2). HSV-1 mainly causes lesions above the waist, such as vesicular stomatitis, herpetic keratitis, herpetic encephalitis, etc. The incidence of this virus infection accounts for about 5% of all encephalitis. [0003] 20%, and the case fatality rate is as high as 70%. Infection during pregnancy not only harms the mother, but also can often cause intrauterine infection and lea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/4375A61P31/22A61P1/02A61P27/02A61P25/00A61P15/00A61P43/00
CPCA61K31/4375A61P31/22A61P1/02A61P27/02A61P25/00A61P15/00A61P43/00
Inventor 曾南唐琼刘鳐饶志粒曾九僧彭利霞王德健
Owner CHENGDU UNIV OF TRADITIONAL CHINESE MEDICINE
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