Synthesis And In Vitro Effect Evaluation Of A New Antiviral Drug KPT-335 And Its Derivatives

Influenza virus is an important pathogen of respiratory diseases.At present,vaccination is mainly used to prevent its infection,but influenza virus is prone to mutation,so the effectiveness of vaccine is facing great challenges.Although Japanese encephalitis can be prevented by vaccine,there are no drugs that can inhibit Japanese encephalitis virus and be used for treatment of Japanese B encephalitis.Therefore,it is very important to develop antiviral drugs that can play a broad-spectrum role in the prevention and treatment of such viral diseases.KPT-335 is a nuclear export protein inhibitor,as a new antiviral drug,can inhibit the proliferation of influenza virus in vivo and in vitro.In this study,KPT-335 was synthesized or modified by chemical methods to generate new derivatives,and then studied its antiviral effect,in order to obtain effective antiviral drugs.1.Synthesis of KPT-335 and its derivatives:Using trifluoromethyl benzonitrile and proparynic acid as initial materials,the final product KPT-335 was successfully synthesized through addition,substitution,hydrolysis and condensation.Based on KPT-335,the corresponding derivatives were synthesized.The structures of the final products and their derivatives were analyzed and identified by 1HNMR and MS.The results showed that the molecular structure and molecular weight of the compound were correct,and 5 new derivatives were successfully synthesized.2.Effects of KPT-335 and its derivatives on influenza virus proliferation.The TCID50 of MDCK cells infected with influenza virus was calculated by Reed-Muench method,and 100 TCID50 cells were determined to be infected.By CCK-8 method of cell survival rate detection,the cell survival rate is more than 80%when the drug concentration is 4 μmol/L,and the cell survival rate is about 60%when the drug concentration is 20 μmol/L.Three drug concentrations of 4 μmol/L,8 μmol/L and 16 μmol/L were selected to verify the inhibitory effect of the drug on influenza virus.After adding the virus venom for incubation for 1 hour,the virus venom was sucked out and the drug KPT-335 and its derivatives were added for further incubation for 48 hours.Under the microscope,the survival cell number of the drug group adding KPT-335 was significantly higher than that of the virus infection group,while the cell survival number of the drug group adding the derivative was significantly reduced.The results showed that KPT-335 could inhibit the proliferation of influenza virus,but the derivative had no significant effect on the proliferation of influenza virus.3.Effects of KPT-335 and its derivatives on the proliferation of Japanese B encephalitis(JE)virus.Taking BHK-21 cells infected by JE virus as a model,the infectivity of JE virus,TCID50 was detected by Reed-Muench method It was determined that 100 TCID50 were used as the inoculation amount for the follow-up test.The toxicity of KPT-335 and its derivatives to BHK-21 cells was determined by CCK-8 assay.The results showed that when the concentration of the drug was 50 μmol/L,the cell survival rate was about 60%.Therefore,three drug concentrations of 5μmol/L,10 μmol/L and 20 lunol/L were selected to investigate the effect of KPT-335 and its derivatives on the proliferation of JE virus.The results showed that KPT-335 had certain inhibitory effect on the proliferation of JE virus,but its derivatives had no significant effect on the virus.The results showed that KPT-335 could inhibit the proliferation of influenza virus and the propagation of JE virus to some extent,but its derivatives had no obvious inhibitory effect on influenza virus and JE virus.Therefore,KPT-335 has a wide application prospect in the prevention and treatment of such viral diseases.