Application of ciprofloxacin in preparation of human cytomegalovirus inhibitor

A technology of human cytomegalovirus and ciprofloxacin, applied in the field of biomedicine

Active Publication Date: 2021-08-20
CANVEST WUHAN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In recent years, research on anti-HCMV drugs has made some progress, mainly based on broad-spectrum antiviral drugs such as ganciclovir, interferon therapy, and symptomatic treatment; A new type of HCMV inhibitor has also shown good antiviral activity in vivo and in vitro tests, but clinical drug-resistant virus strains are also increasing, and there is still an urgent need to develop specific therapeutic drugs against HCMV

Method used

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  • Application of ciprofloxacin in preparation of human cytomegalovirus inhibitor
  • Application of ciprofloxacin in preparation of human cytomegalovirus inhibitor
  • Application of ciprofloxacin in preparation of human cytomegalovirus inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] In this example, after HCMV infects trophoblast cells, it will stimulate the RNAi channel and produce siRNA. The specific process is as follows:

[0024] 1. Experimental materials

[0025] Human trophoblast cells, wild-type HCMV, trophoblast culture medium containing 10% Gibco serum and 2% double antibody, trophoblast medium containing 2% serum, total RNA extraction kit (Gibco).

[0026] 2. Experimental process

[0027] (1) Pave a 24-well plate with trophoblast cells;

[0028] (2) When it grows to 70%-80% confluence, replace the trophoblast culture medium containing 10% Gibco serum and 2% double antibody with the trophoblast medium containing 2% serum (per hole culture medium The amount added is 0.5ml), and 5μl10 was added to each well 7 PFU / ml wild-type HCMV;

[0029] (3) After 48 hours of virus infection, collect samples and extract RNA with a total RNA extraction kit;

[0030] (4) Discard the supernatant, add 350 μl TRK lysate to the well, and put it on the shak...

Embodiment 2

[0042] In this example, a HFF cell model infected with HCMV labeled with green fluorescent protein was established for subsequent research on the inhibitory effect of ciprofloxacin on HCMV. The specific process is as follows:

[0043] 1. Experimental materials

[0044] Human foreskin fibroblasts (HFF), GFP-HCMV labeled with green fluorescent protein, DMEM medium containing 10% Gibco serum and 2% double antibody, DMEM medium containing 2% serum, fluorescence microscope.

[0045] 2. Experimental process

[0046] (1) Utilize the reverse genetics system to construct the GFP-HCMV virus whose genome can express green fluorescent protein;

[0047] (2) Pave a 24-well plate with HFF cells;

[0048] (3) When it grows to 70%-80% confluence, replace the DMEM medium containing 10% Gibco serum and 2% double antibody with the DMEM medium containing 2% serum (the amount of medium added in each well is 0.5ml), add 5μl 10 per well 6 PFU / ml GFP-HCMV;

[0049] (4) After virus infection 4 day...

Embodiment 3

[0052] The present embodiment explores the inhibitory effect of ciprofloxacin (Cipro) on the viral level of HCMV, and the specific process is as follows:

[0053] 1. Experimental materials

[0054] Human foreskin fibroblasts (HFF), GFP-HCMV labeled with green fluorescent protein, DMEM medium containing 10% Gibco serum and 2% double antibody, DMEM medium containing 2% serum, PBS, fluorescence microscope.

[0055] 2. Experimental process

[0056] (1) spread HFF cells to 24-well plate;

[0057] (2) When it grows to 70%-80% confluence, replace the DMEM medium containing 10% Gibco serum and 2% double antibody with the DMEM medium containing 2% serum (the amount of culture medium in each well is 0.5ml), add 5μl 10 per well 7 PFU / ml GFP-HCMV;

[0058] (3) After virus infection 4h, abandon culture medium, wash twice with PBS, add the DMEM culture medium (the addition amount of every well culture medium is 0.5ml) freshly containing 2% serum, simultaneously in experimental group (be...

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Abstract

The invention provides an application of ciprofloxacin in the preparation of human cytomegalovirus inhibitors. By performing small RNA sequencing on HCMV-infected cells, the invention found that after HCMV infection, siRNA will be produced to stimulate the RNAi antiviral pathway, and ciprofloxacin can enhance the RNAi pathway, thereby effectively inhibiting the viral level of HCMV in cells, and It can inhibit the replication level of HCMV virus, so as to prevent and / or treat diseases caused by HCMV infection.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of ciprofloxacin in the preparation of human cytomegalovirus inhibitors. Background technique [0002] Human cytomegalovirus (HCMV) is a DNA virus with the largest genome in the herpesvirus family and belongs to the β-herpesviruses, which can cause lifelong infection in humans. HCMV infection is very common in the population. In developed countries, the positive rate of HCMV antibody in adults is about 50%, while in our country it is as high as 60% to 80%. 10% to 15% of children are first infected with HCMV before the age of 5 years. Primary HCMV infection is generally asymptomatic in healthy hosts but can cause severe and sometimes fatal disease in immunocompromised individuals and neonates. HCMV is the main source of infection leading to congenital malformations in the world. Intrauterine infection of HCMV can cause obvious abnormal symptoms, i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/496A61P31/20
CPCA61K31/496A61P31/20
Inventor 刘愈杰徐国东幸晓莹郑从义
Owner CANVEST WUHAN BIOTECH
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