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Application of Compound ps-341 in the Preparation of Bunyaviridae Phlebovirus Inhibitors

A technology of PS-341 and Bunia virus, applied in the direction of antiviral agents, resistance to vector-borne diseases, dipeptide components, etc., to achieve the effect of inhibiting replication and proliferation

Inactive Publication Date: 2021-08-03
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether the compound PS-341 can inhibit the replication of the Bunyaviridae Phlebovirus virus has not been reported yet.

Method used

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  • Application of Compound ps-341 in the Preparation of Bunyaviridae Phlebovirus Inhibitors
  • Application of Compound ps-341 in the Preparation of Bunyaviridae Phlebovirus Inhibitors
  • Application of Compound ps-341 in the Preparation of Bunyaviridae Phlebovirus Inhibitors

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1, plasmid construction

[0021] Plasmid construction by a method comprising the following steps

[0022] Synthesis of nonstructural protein cDNA of fever with thrombocytopenia syndrome virus (SFTSV), Ukuku virus (UUKV), Sicilian sandfly fever virus (SFSV)

[0023] Retrieve the gene sequences according to the GENBANK gene sequence numbers SFTSV (GI: 747024328), SFSV (GI: 334035), and UUKV (GI: 38371709), and find out the cDNA sequences of SFSV and UUKV respectively. SFTSV, cDNA sequences of SFSV and UUKV.

[0024] Amplification of target gene

[0025] According to the gene sequence SFTSV (GI: 747024328), SFSV (GI: 334035), UUKV (GI: 38371709), primers for viral non-structural proteins (NSs) were designed, the upstream primer restriction site was Sal1, and the downstream primer restriction site was selected For BamH1, an HA tag was added to the end of the downstream primer. Using cDNA as a template, the target gene was amplified according to the PCR instru...

Embodiment 2

[0039] Embodiment 2, cell culture, transfection, activity detection of interferon promoter

[0040] cell culture

[0041] 1) 293T cells ( CRL-3216 TM ) cultivation

[0042] 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.

[0043] 2) HeLa cells ( CCL-2 TM ) cultivation

[0044] 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 UUKV 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) and 200ng VR1012 vector, 50ng SFTSV NSs and 500ng SFSV or UUKV NSs were co-transfected into 293T cells, 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 the application of compound PS-341 in the preparation of Bunyaviridae phlebovirus inhibitors, said compound PS-341 has a structure shown in formula (I): experiments have proved that compound PS-341 can inhibit Bunyaviridae Fever with thrombocytopenia syndrome virus, Ukuku virus, or Sicilian sandfly fever virus in the subviridae Phlebovirus genus. PS‑341 can inhibit the degradation of RIG‑I ubiquitination pathway mediated by SFTSV NSs, so that host cells can activate the interferon antiviral pathway during virus infection, and significantly inhibit virus replication and proliferation.

Description

technical field [0001] The present invention relates to a new application of compound PS-341, in particular to the application of compound PS-341 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 di...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/05A61P31/14
CPCA61K38/05Y02A50/30
Inventor 王涛庞正王志云周振威张仲文
Owner TIANJIN UNIV
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