二氢乳清酸脱氢酶抑制剂ASLAN003抗戊型肝炎病毒的研究

Study on Anti-Hepatitis E Virus Effect of Dihydroorotate Dehydrogenase Inhibitor ASLAN003

研究二氢乳清酸脱氢酶抑制剂ASLAN003对戊型肝炎病毒(HEV)复制的影响,为抗HEV治疗提供新型候选药物。用带有高斯荧光素酶的HEV复制子模型研究ASLAN003是否具有抗HEV作用,实时荧光定量PCR和间接免疫荧光检测ASLAN003对HEV RNA复制以及衣壳蛋白表达的抑制作用,细胞毒性试验和间接免疫荧光计算ASLAN003的半数毒性浓度(CC_(50))和半数抑制浓度(IC_(50))。结果表明,ASLAN003在0.2~50μmol/L浓度范围内能显著抑制HEV复制子在HEK 293T和Huh7细胞内的复制;ASLAN003在1μmol/L浓度能有效抑制HEV RNA复制,使其下降50%;在0.4μmol/L的浓度能够显著抑制衣壳蛋白ORF2的表达。通过对ASLAN003的细胞毒性和抑制HEV效果进行研究发现,其CC_(50)为294μmol/L,而IC_(50)为0.231μmol/L。说明ASLAN003能够有效抑制HEV复制,提示其可作为潜在抗病毒药物。

The aim of the study is to investigate the effect of dihydroorotate dehydrogenase inhibitor ASLAN003 on Hepatitis E virus,which will provide a potential novel drug candidate for combating HEV infection.The antiviral effect of ASLAN003 on HEV was investigated by using HEV Gaussia luciferase replicon model,real-time quantitative PCR and indirect immunofluorescent assay were used to detect the effects of viral replication and the expression level of HEV capsid protein,the half-toxicity concentration(CC_(50))and half-inhibitory concentration(IC_(50))of ASLAN003 were determined by cell toxicity assay and indirect immunofluorescent assay.The results showed that:ASLAN003 significantly inhibited HEV replication with the drug concentration ranging from 0.2μmol/L to 50μmol/L.In HEV infectious cell models,both the levels of HEV RNA and capsid protein ORF2 were significantly suppressed upon ASLAN003 treatment.Specifically,1μmol/L of ASLAN003 inhibited HEV RNA replication by 50%,and 0.4μmol/L of ASLAN003 efficiently reduced the expression of capsid protein ORF2.The CC_(50) of ASLAN003 is 294μmol/L,whereas the IC_(50) is 0.231μmol/L.ASLAN003 possesses the efficacy to potently inhibit HEV,severing as a potential antiviral candidate for combating HEV infection.