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New Application of Corydalin in Prevention and Treatment of Human Cytomegalovirus Infection

A technology of human cytomegalovirus and Corydalin, applied in the field of biomedicine, can solve problems such as disorder, drug resistance, bone marrow suppression, etc.

Active Publication Date: 2021-12-14
ZHEJIANG HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no approved vaccine on the market, and the clinical symptomatic drugs are mainly antiviral drugs such as foscarnet sodium, cidofovir, and fomivirsen, but these drugs have obvious side effects such as bone marrow suppression, nephrotoxicity, and electrolyte imbalance. And there will be drug resistance, which limits its clinical application
[0003] Corydalis is a kind of alkaloid derived from Corydalis buchneri of the Papaveraceae plant, so far, there is no application of corydalin in the preparation of drugs for the prevention and / or treatment of HCMV

Method used

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  • New Application of Corydalin in Prevention and Treatment of Human Cytomegalovirus Infection
  • New Application of Corydalin in Prevention and Treatment of Human Cytomegalovirus Infection
  • New Application of Corydalin in Prevention and Treatment of Human Cytomegalovirus Infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Corydalin alone treatment on the influence of WI-38 cell proliferation activity

[0062] The cell viability was detected by the MTT method as follows: PD30 human embryonic lung diploid fibroblast WI-38 was seeded in a 96-well cell culture plate at a cell density of 3000 per well, and after 24 hours of culture, different concentrations of Corydalis were added For each concentration, 3 parallel wells were set up, and a blank control group without drugs and a solvent control group without cells were set up. After culturing in the incubator for 48 hours, add 20 μl of MTT (5 mg / ml, prepared with serum-free DMEM medium) to each well, at 37°C, 5% CO 2 Continue culturing for 4 hours under the same conditions, then aspirate the liquid in the cell culture wells, add 150 μl DMSO to each well, and shake gently on the shaker for 10 minutes to fully dissolve the crystals, then measure the light absorbance at 570 nm, and calculate the relative cell viability. Take the cell ...

Embodiment 2

[0063] Embodiment 2 Corydalin treatment and the inoculation of HCMV

[0064] Human embryonic lung fibroblast WI-38 cells using PD30, with medium containing 10% FBS according to 2 × 10 4 / cm 2The amount of cells was plated on a six-well cell culture plate, and the medium of 0.2% FBS was replaced after 24 hours to continue culturing for 48 hours. By means of serum starvation, the cell cycle G0 / G1 was synchronized, which was beneficial to the infection of HCMV. Afterwards, the HCMV virus (Towne virus strain) was inoculated with an inoculation dose of 0.01 MOI (multiplicity of infection), and the culture was continued until the specified time for relevant detection. When testing the anti-HCMV activity of Corydalin, a certain concentration of Corydalin was added to the medium 2 hours in advance, and then HCMV was inoculated (MOI0.01), and then changes in viral protein expression and DNA amplification were observed.

Embodiment 3

[0065] Embodiment 3Western-blot detects the impact of Corydalin on the expression of HCMV virus protein

[0066] In order to confirm the anti-HCMV effect of cordoresin, we detected the effect of cordoresin on the expression of HCMV immediate early protein IE1 / 2 and early protein UL44, cell culture and cordoresin treatment and HCMV inoculation as in Example 2 above. Set up a control group without any drug treatment and only inoculated HCMV; solvent DMSO treatment, a control group inoculated with HCMV, a drug control group inoculated with HCMV and clinically treated with foscarnet sodium (200 μg / ml), and Corydalin (10 μM) treatment Experimental groups with different time of inoculation with HCMV. 1, 4, and 7 days after HCMV inoculation, the supernatant was discarded, the cells were washed three times with PBS, the cells were lysed with medium-strength RIPA lysate, and the cell samples were collected. After the protein concentration was determined with the BCA kit, the correspond...

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Abstract

The invention belongs to the field of biotechnology, and the invention provides a new application of Corydalis line in the preparation of medicines for preventing and treating human cytomegalovirus infection. Experiments show that Corydalin has obvious anti-HCMV effect. Corydalin (1‑20 μM) alone treated human cytomegalovirus HCMV host cell-human embryonic lung diploid fibroblast WI‑38 did not show obvious cytotoxicity, and cordylyl could significantly improve the cytopathy formed by HCMV infection effect, inhibit the expression of HCMV characteristic immediate early protein IE1 / 2 and early protein UL44, effectively inhibit the DNA replication of HCMV, and have the application prospect of preparing anti-HCMV drugs.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a new application of Corydalis line in preventing and treating human cytomegalovirus infection. Background technique [0002] Human cytomegalovirus (HCMV) belongs to the β-subfamily of herpesviruses, and the population is generally susceptible to it, and is infected for life after infection. After the infection is relieved, the virus stays latent in the body for a long time, forming a latent infection. In transplant patients, secondary infection is prone to occur, which can easily lead to damage to the nervous system, liver, respiratory system and blood system, and even life-threatening in severe cases; at the same time, in pregnant women, secondary infection of HCMV is the cause of congenital diseases in newborns The main pathogens cause jaundice hepatitis, mental retardation, blindness, deafness and other multi-organ and multi-system lesions. Therefore, active and effective treatmen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/58A61K36/66A61P11/00A61P31/22
CPCA61K31/58A61K36/66A61P11/00A61P31/22
Inventor 毛根祥王三应徐小刚万晓青苏慧丽张婧孙川代继桓邢文敏
Owner ZHEJIANG HOSPITAL
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