Application of Z-VAD-FMK in preparation of human herpes simplex virus type 1 inhibitor

The technology of Z-VAD-FMK and 1.Z-VAD-FMK is applied in the application field of Z-VAD-FMK in the preparation of human herpes simplex virus type 1 inhibitor, and can solve the problem that there is no effective vaccine and medicine for HSV-1 And other issues

Active Publication Date: 2020-11-17
CANVEST WUHAN BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

HSV-1 has become an important factor affecting people's life and health, and there is currently no effective vaccine or drug against HSV-1

Method used

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  • Application of Z-VAD-FMK in preparation of human herpes simplex virus type 1 inhibitor
  • Application of Z-VAD-FMK in preparation of human herpes simplex virus type 1 inhibitor
  • Application of Z-VAD-FMK in preparation of human herpes simplex virus type 1 inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] This example explores the hydrolysis of Caspase 3 on the key proteins p65, RelB, and C-Rel in the NF-κB signaling pathway. The specific process is as follows:

[0027] 1. Experimental materials

[0028] 293T cells, MEM medium containing 10% Gibco serum and 2% double antibody, transfection reagent, MEM medium containing 2% serum, plasmid expressing HA-tagged Caspase 3 and Caspase 3 activity loss (Caspase 3mut), Plasmid expressing IL-1β, protein lysate, p65 endogenous antibody, RelB endogenous antibody, C-Rel endogenous antibody, HA antibody, Tubulin antibody, western bolt related reagents.

[0029] 2. Experimental process

[0030] (1) Spread 6-well plates with 293T cells;

[0031] (2) When it grows to 70%-80% confluence, replace the MEM medium containing 10% Gibco serum and 2% double antibody with the MEM medium containing 2% serum (the amount of culture medium in each well is 1ml);

[0032] (3) Using a transfection reagent, transfer 1 μg of empty-load, Caspase 3, an...

Embodiment 2

[0045] This example explores the hydrolysis of Caspase 3 on the key protein IRF3 in the IFN-I signaling pathway. The specific process is as follows:

[0046] 1. Experimental materials

[0047] 293T cells, MEM medium containing 10% Gibco serum and 2% double antibody, transfection reagent, MEM medium containing 2% serum, plasmid expressing HA-tagged Caspase 3 and Caspase 3 activity loss (Caspase 3mut), Plasmid expressing Flag-tagged IRF3 (Flag-IRF3), protein lysate, Flag antibody, HA antibody, Tubulin antibody, westernbolt-related reagents.

[0048] 2. Experimental process

[0049] (1) Spread 6-well plates with 293T cells;

[0050] (2) When it grows to 70%-80% confluence, replace the MEM medium containing 10% Gibco serum and 2% double antibody with the MEM medium containing 2% serum (the amount of culture medium in each well is 1ml);

[0051] (3) Using a transfection reagent, transfer 1 μg of empty load, Caspase 3, Caspase 3mut, and Flag-IRF3 plasmids into 293T cells in a co...

Embodiment 3

[0063] This embodiment explores the influence of Caspase 3 on the mRNA level of CXCL10 stimulated by HSV-1, and the specific process is as follows:

[0064] 1. Experimental materials

[0065] Mouse macrophages (Macrophages), DMEM medium containing 10% Gibco serum and 2% double antibody, Caspase 3 inhibitor, DMEM medium containing 2% serum, HSV-1 virus, total RNA extraction kit (Gibco ), onestep qRT-PCR kit.

[0066] 2. Experimental process

[0067] (1) Pave a 24-well plate with mouse macrophages;

[0068] (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 6 PFU / mlHSV-1 virus;

[0069] (3) After 4 hours of virus infection, the supernatant was discarded, and washed twice with PBS, and Caspase3 inhibitor was added to the corresponding wells, and the samples were collected after 24 hours of virus infection an...

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Abstract

The invention provides an application of Z-VAD-FMK in preparation of a human herpes simplex virus type 1 inhibitor. According to the application, cell apoptosis is inhibited through Z-VAD-FMK, and NF-kB and IFN-1 signal pathways excited in different modes can be inhibited by cell apoptosis no matter at a cell level or an animal level, so that natural immune response caused by human herpes simplexvirus type 1 is negatively regulated and controlled by cell apoptosis; and therefore, the Z-VAD-FMK is used for enhancing a natural immune pathway excited by the human herpes simplex virus type 1, andthe purpose of effectively resisting viruses is achieved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of Z-VAD-FMK in the preparation of human type 1 herpes simplex virus inhibitors. Background technique [0002] Herpes virus (Herpes virus) is a large class of encapsulated dsDNA virus, according to its biological characteristics, classified as Herpesviridae (Herpesviridae). More than 100 kinds of herpes viruses have been found so far. Herpes simplex virus (HSV) is a typical representative of herpes viruses. It mainly invades skin, mucous membranes and nerve tissues, seriously affecting human life and health. [0003] HSV belongs to the herpesvirus subfamily Alphaherpesvirinae (Alphaherpesvirinae) and the genus Simplexvirus. HSV viruses can be divided into: human herpes simplex virus type 1 (HSV-1) and human herpes simplex virus type 2 (HSV -2). HSV-1 mainly infects mucosal epithelial cells and establishes lifelong infection in sensory neurons. Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/06A61P31/22
CPCA61K38/06A61P31/22
Inventor 徐国东刘愈杰幸晓莹郑从义
Owner CANVEST WUHAN BIOTECH
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