Novel applications of histone deacetylase inhibitor in treatment of beta subfamily herpesvirus

A sirtuin and herpes virus technology, applied in antiviral agents, scientific instruments, biochemical equipment and methods, etc., can solve the problems of no effective treatment and great harm

Inactive Publication Date: 2016-06-01
INST PASTEUR OF SHANGHAI CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Human cytomegalovirus is the most common and harmful virus in in

Method used

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  • Novel applications of histone deacetylase inhibitor in treatment of beta subfamily herpesvirus
  • Novel applications of histone deacetylase inhibitor in treatment of beta subfamily herpesvirus
  • Novel applications of histone deacetylase inhibitor in treatment of beta subfamily herpesvirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1 Pre-dissolution storage of vorinostat (SAHA)

[0083] Preparation and storage of the mother liquor of vorinostat: accurately weigh vorinostat and dissolve it in DMSO at a concentration of 10mM, take a 1.5mL centrifuge tube, aliquot 20μL per tube, and store at -20℃.

Embodiment 2

[0084] Example 2 Titer experiment to determine the relationship between the concentration of vorinostat and its antiviral ability

[0085] The preparation of the medium: 500mLDMEM, 50mLFBS, 500μL antibiotic P / S, and mix well.

[0086] The above medium is used to culture HF cells, and the cells are cultured in 10cm petri dishes, and each dish uses 10 mL of medium each time. Culture HF cells in a 10cm petri dish, and when they are full, digest them with 1mL trypsin. If you want to divide it into a 12-well plate, add it to 15mL of medium and mix well, divide 1mL per well into each well, and place in 5% CO 2 , 37 ℃ constant temperature incubator for 40 hours, it can be used for virus infection; if it is to be divided into 96-well plates, add to 30mL medium and mix well, divide 100μL per well into each well, and place in 5% CO 2 , Incubate in a constant temperature incubator at 37°C for 40 hours, which can be used for virus titer testing.

[0087] Dilution of vorinostat: According to the...

Embodiment 3

[0101] Example 3 Fluorescence quantitative PCR was used to detect the effect of 5 μM vorinostat on HCMV virus DNA replication.

[0102] Preparation of 2×digestion buffer: NaCl 1.1688g, Tris0.24228g, EDTA 1.8612g, SDS1g, add sterile water to 100mL, mix well, and store at room temperature.

[0103] The preparation of 7.5M ammonium acetate: In a clean 250ml beaker, first add about 30ml of sterile water, weigh 57.81g of ammonium acetate, mix well with a magnetic stirrer, dilute to 100ml, filter with 0.22μm filter into two 50ml centrifuge tubes , Store at room temperature.

[0104] Extraction steps of cell DNA and virus DNA:

[0105] (1) Separate the cells in a 12-well plate as described in Example 2. After culturing for 40 hours, they were infected with WT-GFP virus, and treated with vorinostat at the same time. The samples were collected after a certain period of time after infection.

[0106] (2) After the time is up, collect the samples from the corresponding wells, and wash each well w...

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PUM

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Abstract

The invention discloses novel applications of a histone deacetylase inhibitor in treatment of beta subfamily herpesvirus, and more specifically provides applications of a histone deacetylase (HDAC) inhibitor or pharmaceutically acceptable salts in preparing of pharmaceutical compositions used for treating beta subfamily herpesvirus. It is confirmed by experiments that low concentration HDAC inhibitor possesses relatively high inhibition efficiency on beta subfamily herpesvirus infection, and inhibition rate is as high as 99.69 to 99.97%. So that the HDAC inhibitor can be taken as an effective drug used for treating beta subfamily herpesvirus infection in clinical.

Description

Technical field [0001] The present invention relates to the field of virus therapy. Specifically, it relates to a new use of a histone deacetylase inhibitor to treat β subfamily herpes virus. Background technique [0002] Herpesviruses are a group of medium-sized double-stranded DNA viruses, which are divided into three subfamilies: α, β, and γ according to their physical and chemical properties. Alpha herpes viruses (such as herpes simplex virus, varicella-zoster virus) multiply fast and cause cell pathology. Beta herpes virus (such as cytomegalovirus) has a long growth cycle, and infected cells form giant cells. γ herpes virus (such as Epstein-Barr virus), the target cell of infection is lymphoid cells, which can cause lymphatic hyperplasia. The herpes virus infection has a wide host range, which can infect humans and other vertebrates. There are 7 kinds of herpes viruses that cause humans to produce. [0003] Human cytomegalovirus (HCMV) belongs to the β subfamily of the he...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K31/47A61K31/167A61K31/42A61P31/22G01N21/64C12Q1/70C12Q1/68G01N33/569
Inventor 钱志康宣宝琴刘中顺
Owner INST PASTEUR OF SHANGHAI CHINESE ACADEMY OF SCI
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