Application of compound MLN4924 in preparation of bunyaviridae phlebovirus virus inhibitor

A bunya virus, virus inhibitor technology, used in antiviral agents, resistance to vector-borne diseases, medical preparations containing active ingredients, etc.

Inactive Publication Date: 2018-06-29
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Previously, research reports on MLN4924 mostly focused on its anti-tumor properties by blocking the cell cycle or causing apoptosis, and there has been no report on its anti-viral effect and its mechanism.

Method used

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  • Application of compound MLN4924 in preparation of bunyaviridae phlebovirus virus inhibitor
  • Application of compound MLN4924 in preparation of bunyaviridae phlebovirus virus inhibitor
  • Application of compound MLN4924 in preparation of bunyaviridae phlebovirus virus inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1, plasmid construction

[0020] Plasmid construction by a method comprising the following steps

[0021] Synthesis of Nonstructural Protein cDNA of Fever with Thrombocytopenia Syndrome Virus (SFTSV) and Sicilian Sandfly Fever Virus (SFSV)

[0022] According to GENBANK gene sequence numbers SFTSV (GI: 747024328) and SFSV (GI: 334035), the gene sequence was retrieved, and the cDNA sequences of SFTSV and SFSV non-structural protein NSs were synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd.

[0023] Amplification of target gene

[0024] According to the sequence of SFTSV and SFSV non-structural protein NSs genes, NSs-specific primers were designed, the restriction site of the upstream primer was Sal1, the restriction site of the downstream primer was BamH1, and an HA tag was added at the end of the downstream primer. Using cDNA as a template, the target gene was amplified according to the PCR instruction system.

[0025]

[0026] PCR product gel reco...

Embodiment 2

[0038] Embodiment 2, cell culture, transfection, activity detection of interferon promoter

[0039] cell culture

[0040] 1) 293T cells ( CRL-3216 TM ) cultivation

[0041] 293T cells were cultured in DMEM liquid medium containing 10% FBS, 1% penicillin-streptomycin, placed in an incubator containing 5% CO2 at 37°C, and incubated in an incubator containing 0.25% EDTA after overgrowth. Enzyme digestion, after about 2-3min, add 10% FBS fresh medium to terminate, centrifuge at room temperature at 1200rpm for 3min, discard the supernatant, take an appropriate amount of fresh medium to resuspend, return one-third of the culture medium to the culture bottle, and the remaining Transfer to 24-well cell culture plates.

[0042] 2) HeLa cells ( CCL-2 TM ) cultivation

[0043] HeLa cells were cultured in DMEM liquid medium containing 10% FBS and 1% penicillin-streptomycin, and incubated at 37°C in an incubator containing 5% CO2. After about 2-3 days, digest with trypsin contain...

Embodiment 3

[0059] Example 3, SFTSV SFSV NSs inhibit IFN-β promoter activity

[0060] Perform experiments such as cell transfection, fluorescent dual reporter analysis, and Western blot according to the specified steps (the method is the same as in Example 2)

[0061] 200ng RIG-I-Flag plasmid (Addgene) was co-transfected into 293T cells with 200ng VR1012 vector, 50ng SFTSV NSs and 500ng SFSV or NSs respectively, and 500ng IFN-β-Luc and 50ng pRL-TK plasmids were transfected at the same time. Forty-eight hours after transfection, the expression of the reporter gene was detected by a dual reporter gene detection method. NSs protein expression level by Western-Blot results as follows figure 1 show. The results of dual reporter gene detection showed that the NSs protein of the virus significantly inhibited the activation of the RIG-I-induced interferon beta promoter, which also indicated that the NSs protein inhibited the production of host type I interferon.

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Abstract

The invention discloses an application of a compound MLN4924 in preparation of a bunyaviridae phlebovirus virus inhibitor. The compound MLN4924 has a structure shown in the formula (I) (shown in the specification), and experiments prove that the compound MLN4924 can inhibit severe fever with thrombocytopenia syndrome viruses or Sicilian phlebotomus fever viruses in bunyaviridae phlebovirus viruses. According to the invention, ubiquitination degradation of RIG-I mediated by SFTSV NSs is inhibited, so that a host cell activates an interferon antiviral pathway during virus infection, and replication and proliferation of viruses are obviously inhibited.

Description

technical field [0001] The present invention relates to a new application of compound MLN4924, in particular to the application of compound MLN4924 in the preparation of Bunyaviridae phlebovirus inhibitors. Background technique [0002] In recent years, due to factors such as global climate change, several infectious diseases that have caused great harm to human health have occurred in my country, such as hand, foot and mouth disease, bird flu, SARS, and streptococcal suis. These highly infectious diseases have posed serious challenges to our country's society, economy, and national security. Scientific research on effective prevention methods for these high-risk pathogens is of certain importance and urgency. Research on specific therapeutic drugs and vaccines for emerging infectious diseases is crucial to the prevention and control of infectious diseases in my country. [0003] Since 2009, major media have continuously reported that tick bites have caused the epidemic of ...

Claims

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Application Information

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IPC IPC(8): A61K31/519A61P31/14
CPCA61K31/519Y02A50/30
Inventor 王志云王涛庞正李雪平潘明磊
Owner TIANJIN UNIV
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