| Viral infections lead to the occurrence and spread of a large number of infectious diseases,causing huge global economic losses.Vaccination is an effective way to prevent virus infection,but the mutant strain of the virus greatly reduces the protective efficiency of the vaccine.There is great potential for small molecular drugs to treat viral diseases,compounds containing triazine or pyrimidine ring show broad-spectrum biological activities,such as antiviral,antibacterial,anticancer and anti-inflammatory.PIKfyve is a phosphoinositol kinase containing FYVE fingers,reported as a potential antiviral target,and its inhibitors are mostly triazine or pyrimidine derivatives.In this study,1,3,5-triazine or pyrimidine derivatives were synthesized and their broad-spectrum antiviral effects were preliminarily evaluated.In this study,a series of derivatives with morpholine substituted 1,3,5-triazine or pyrimidine ring as parent rings were designed,and thirty compounds were obtained.Twenty-six 1,3,5-triazine derivatives were obtained by nucleophilic substitution and condensation reaction using cyanogen chloride as raw material,among which 10 bc,10bd,10 ca,10ca,10 cb,10cc,10 cd,10db,10 dc and 10 dd were synthesized for the first time.The effects of alkaline conditions,solvents,temperature and reaction time on the yield were investigated.The overall yield was between 48.82(4)and 71.11(4);Using compound 15 as raw material through nucleophilic substitution and condensation reaction,four kinds of pyrimidine derivatives were obtained.The acid condition and high temperature can lead to the rupture of ether bond in the product.The optimization of reaction conditions were carried out,and the total yield is up to86.65(4).All obtained fifty-three compounds,including twenty-three intermediates,were characterized by1 H NMR,13 C NMR and HRMS.The effect of thirty compounds on the activity of PK-15 cells was detected by CCK-8 kit,and the results were shown as CC50.The effect of these compounds on the proliferation of PRV-GFP and VSV-GFP was determined by flow cytometry,and the results were shown as IC50.SI was used to evaluate the drug inhibition rate of compounds comprehensively,and all three effective compounds including 10 ce,10de and 16 c were selected.TCID50 was used to measure progeny virus titers.WB was used to detect the protein expressions of PRV g B and VSV GFP.q RT-PCR was used to detect the m RNA transcribe of PRV-g B and VSV-N to evaluate the antiviral activity of the compounds.The results showed that compounds 10 ce,10de and 16 c inhibited the replication of progeny viruses,the expression of PRV g B and VSV GFP proteins,and the m RNA transcribe of PRV-g B and VSV-N in PK-15 cell species.Finally,molecular docking was used to predict the docking status of the compounds with PIKfyve.The results showed that compounds 10 de and 16 c can interact with ASP-375 and MET-384 through hydrogen bonding and occupy the cavity of PIKfyve protein well.In conclusion,the results of this study not only enriched the kinds of1,3,5-triazine or pyrimidine derivatives,but also screened out three compounds with good antiviral activity.The broad-spectrum antiviral activity was preliminarily demonstrated and compound 10 de was predicted to be a potential inhibitor of PIKfyve,which provided a new strategy for the development and research of antiviral drugs. |
