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Alkaloid for resisting herpes virus infection and combination and application

An anti-herpes virus, herpes simplex virus technology, applied in the direction of antiviral agents, medical preparations containing active ingredients, organic active ingredients, etc. The problem of high incidence of reaction, to achieve the effect of regulating immune inflammatory response

Inactive Publication Date: 2020-11-27
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in recent years, there have been more and more reports about VZV resistance to ACV and / or PFA and the poor prognosis of patients infected with drug-resistant strains. Acyclovir antiviral combined with analgesic drugs are often used in clinical treatment of herpes zoster, and the incidence of adverse reactions is high. Risk of acute kidney injury
Acyclovir only has anti-VZV activity, but cannot alleviate the immune inflammatory response and neuropathic pain caused by the virus

Method used

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  • Alkaloid for resisting herpes virus infection and combination and application
  • Alkaloid for resisting herpes virus infection and combination and application
  • Alkaloid for resisting herpes virus infection and combination and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: Benzoyl aconitine, benzoyl hypoaconitine, benzoyl neoaconitine, aconitine, hypoaconitine, neoaconitine have good anti-herpes simplex in vitro Viral activity.

[0036] The CPE method detects the inhibitory effect of drugs on HSV-1 and HSV-2: culture Vero cells in good growth state to monolayer, and use 100TCID 50 The HSV-1 and HSV-2 virus liquids were adsorbed and inoculated for 2 hours, then the supernatant was discarded, and 7 concentrations (10-1000 μM) of benzoyl aconitine, benzoyl aconitine, and benzoyl aconitine were added to each well. Acyl aconitine, aconitine, hypoaconitine, neoaconitine, 5 replicate wells for each concentration, set up cell control, virus control and positive drug (ACV) control, at 37 ° C, 5% CO 2 The culture was continued in the incubator, and the CPE of the cells was observed daily. When the cell CPE of the virus control group reached more than 90%, the cell CPE situation of each well was recorded, and the inhibitory concentr...

Embodiment 2

[0038] Example 2: Benzoylaconitine inhibits the expression of HSV-1 genes UL12, UL42, and UL54 RT-qPCR detection of HSV-1 related gene expression: 100 TCID 50 HSV-1 infected monolayer Vero cells for 2 hours, discarded the supernatant, added 40, 80, and 160 μM benzoylaconitine to the culture wells, and set virus control at the same time, 37 ° C, 5% CO 2 After 16 h in the incubator, total RNA was extracted from the cells. The extracted total RNA was used as a template and reverse-transcribed into cDNA for RT-qPCR experiments. The RT-qPCR reaction system was 5 μL of 2×SYBRGreen PCR MasterMix, 0.5 μL of upstream and downstream primers, and 4 μL of 10-fold diluted cDNA template. PCR reaction conditions: pre-denaturation at 95°C for 2min, denaturation at 95°C for 5s, annealing / extension at 60°C for 10s, a total of 45 cycles. The results were analyzed using CFX Manager 96 software.

[0039] Test results: HSV-1 replication-related gene amplification results Use the cDNA reverse-tr...

Embodiment 3

[0040] Embodiment 3: Benzoyl aconitine inhibits the mRNA expression of HSV-1 glycoprotein gD. Nucleic acid molecular hybridization detects the gene expression of HSV-1 glycoprotein gD: According to the HSV-1 complete gene sequence in GenBank, design corresponding to HSV- 1 An oligonucleotide probe of the glycoprotein gD gene sequence, the probe sequence was synthesized by Shanghai Sangong Biotechnology Company. Vero cells made 2 x 10 5 / ml cell suspension, inoculated in culture flasks with coverslips, added 1.8ml cells to each well, and placed in 5% CO 2 Incubate in an incubator at 37°C for 24h-48h, discard the growth medium, and add about 100TCID 50 100 μl of virus solution, adsorb for 2 hours at 37°C, suck out the virus solution, add 200 μl of PSP diluted with maintenance solution, place in 5% CO 2Cultured at 37°C in an incubator. After 24 hours, take out the cell fly piece, dehydrate with gradient ethanol, rinse twice with PBS, digest with 1 μg / ml proteinase K at 37°C fo...

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Abstract

The invention provides an alkaloid for resisting herpes virus infection. The alkaloid comprises one or more of the following alkaloids: benzoyl aconitine (C32H45NO10), benzoyl hypacoitine (C31H43NO9),benzoyl mesaconine (C31H43NO10), aconitine (C34H47NO11), hypacoitine (C33H45NO10), and mesaconine (C33H45NO11), as well as a composition containing the alkaloid, wherein the alkaloid and the composition thereof have good anti-herpes virus infection effects. The alkaloid and the composition thereof are applied to preparation of drugs for preventing or treating herpes simplex virus type 1 or herpessimplex virus type 2 infection or varicella-zoster virus infection.

Description

technical field [0001] The present invention relates to the field of biotechnology. Background technique [0002] Herpes simplex virus is a linear double-stranded DNA virus. Although it can cause experimental animal infection, humans are its only natural host. Herpes simplex virus (HSV) is divided into two serotypes according to the physical and chemical properties, biological characteristics, DNA restriction endonuclease map and temperature sensitivity of the virus: herpes simplex virus type 1 (HSV-1) and simplex virus (HSV-1). Herpes virus type 2 (HSV-2), there are common antigens and specific antigens between the two types. The nucleotide sequence homology between the two types reaches 47%-50%, and the genome structure is basically the same. The HSV DNA molecule is about 150kb in length and has more than 70 coding genes, which are presumed to encode at least 70 different viral proteins. HSV-1 mainly causes various diseases such as peripheral facial paralysis, facial he...

Claims

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Application Information

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IPC IPC(8): A61K31/439A61P31/22
CPCA61K31/439A61P31/22A61K2300/00
Inventor 荆凡波周长凯纪洪艳邢晓敏
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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