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Application of ALKBH5 inhibitor in treatment of virus infectious diseases

A technology related to infectious diseases and viral infections, applied in the fields of biotechnology and medicine, can solve problems affecting virus replication and infection

Active Publication Date: 2019-11-08
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, what role ALKBH5 plays in the process of virus infection and whether it affects virus replication and infection by regulating the production of natural immune factor interferon remains to be further explored.
[0009] At present, there is no related RNA m 6 A research report on the regulation mechanism of host response to virus infection and the therapeutic application of A demethylase ALKBH5 in virus-related diseases

Method used

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  • Application of ALKBH5 inhibitor in treatment of virus infectious diseases
  • Application of ALKBH5 inhibitor in treatment of virus infectious diseases
  • Application of ALKBH5 inhibitor in treatment of virus infectious diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0119] Example 1: ALKBH5 promotes viral infection in mouse macrophages

[0120] A: Knockdown of ALKBH5 in mouse macrophages using interfering RNA (siRNA) can inhibit virus replication

[0121] The primary peritoneal macrophages of mice were isolated from C57BL / 6 small cells after intraperitoneal injection of 2ml of 3% thioglycolate (according to % is the ratio of m / v, dissolved in double distilled water, and then subjected to high-pressure sterilization). Rats (6-8 weeks old, male) were cultured in endotoxin-free high-glucose DMEM medium containing 10% fetal bovine serum (FBS).

[0122] The siRNA used to interfere with the mouse ALKBH5 gene was transfected into primary peritoneal macrophages using TransIT-siQUEST Transfection Reagent (purchased from Mirus Company) according to the operating instructions (transfection parameters for 12-well cell culture plates: no 100 μl of serum medium, 3 μl of TransIT-siQUEST Transfection Reagent, 2.8 μl of siRNA at a final concentration o...

Embodiment 2

[0138] Example 2: Effects of ALKBH5 gene knockout on the survival rate of virus-infected mice and the virus content in the body

[0139] ALKBH5 knockout mice (ALKBH5KO) were obtained from the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences (Zheng G et al., Mol Cell, 2013, 49(1):18-29), and had been crossed with mice of C57BL / 6 background10 For more than one generation, the genetic background is guaranteed to be C57BL / 6. Take 6 to 8 weeks old healthy littermates of wild-type and gene-deleted mice of the same sex for the experiment, and conduct the experiment according to 5×10 7 The ratio of PFU / g body weight was carried out by intravenous injection for virus infection in vivo, and then the survival period, virus titer detection of major organs, and corresponding tests for lung histochemical detection were carried out.

[0140] By observing and counting the survival period of mice infected with the above doses of virus, the survival rate of mice ( fig...

Embodiment 3

[0143] Example 3: ALKBH5 promotes the replication of different types of RNA viruses and DNA viruses in mice

[0144] The sources and usage methods of RNA virus vesicular stomatitis virus (Vesicular Stomatitis Virus, VSV) and DNA virus herpes virus (Herpes simplex virus type 1, HSV-1) were in accordance with previous literature reports (Wang P et al., Science, 2017, 358( 6366):1051-1055; Liu S et al., Nature immunology, 2018, 19(1):41-52). The wild-type and ALKBH5 gene-deficient mice of the same sex in the same sex at the age of 6 to 8 weeks were used for the experiment, and the RNA virus VSV in vivo was injected intravenously (according to 5×10 7 The ratio of PFU / g body weight) or DNA virus HSV-1 (according to 1×10 7 PFU / mouse) infection, 18 hours later, blood was taken from the eyeball, and then the liver, spleen, lung or brain tissue of the mouse was taken out. RNA in cells was extracted by Trizol method and reverse-transcribed, and real-time fluorescent quantitative PCR (...

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Abstract

The invention relates to application of an AlkB homologous 5 (ALKBH5) inhibitor of demethylase in the treatment of virus infectious diseases. In particular, the invention relates to the use of the ALKBH5 inhibitor in the preparation of products for the treatment of the virus infectious diseases and / or diseases and / or symptoms related to virus infection, and a corresponding pharmaceutical composition or kit of the products. The application can be used for inhibiting and treating the virus infection, especially for objects with abnormal natural immune functions, and has a wide application prospect.

Description

technical field [0001] This disclosure is in the fields of biotechnology and medicine. Specifically, the present disclosure relates to the application of inhibitors of demethylase AlkB homolog 5 (AlkB homolog 5, RNA demethylase, ALKBH5) in the treatment of viral infectious diseases. Background technique [0002] Viral infection can cause various diseases and threaten human health. Studying the mechanism of virus-host interaction will help to understand the process of viral infection and its regulatory molecular mechanism, and provide new drug targets for the prevention and treatment of viral infectious diseases. [0003] Innate immune receptors can activate the kinase TBK1 and transcription factor IRF3 in the downstream signaling pathway by recognizing the virus that infects the host cell, thereby inducing the body to produce interferon (Interferon, IFN) and activating the expression of antiviral effector genes to resist infection with the virus. Virus clearance (Schneider...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/06A61K38/16A61K48/00A61P31/14A61P31/20A61P31/12C12Q1/70C12Q1/02C12R1/93
CPCA61K38/16A61K45/06A61K48/005A61P31/12A61P31/14A61P31/20C12Q1/025G01N21/6486G01N33/5008G01N33/5023A61K2300/00
Inventor 刘洋曹雪涛
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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