人干扰素α1b体外抗呼吸道病毒的药效学研究

摘要/Abstract

摘要： 目的：评价人干扰素（interferon，IFN）α1b体外广谱抗呼吸道病毒的药效。方法：采用CPE（细胞病变）法检测IFNα1b和阳性对照药对MDCK、U937、LLC-MK2、Hep-2、Vero E6、BHK-21 、RD、A549细胞的毒性；采用神经氨酸酶活性检测法检测IFNα1b对流感病毒株CIV-H1N1/CNIC-1909(H1N1)、A/kansas/14/2017(NYMCX-327)(H3N2)和B/Maryland/15/2016（NYMCBX-69A)的抑制效果；采用ELISA试剂盒检测法检测IFNα1b对A型流感病毒株（H1N1，A/PuertoRico/8/1934）及引起的炎症因子的抑制效果；采用间接免疫荧光检测法(IFA)检测IFNα1b对呼吸道合胞病毒（RSV A2）的抑制效果以及对人副流感病毒（HPIV3）的抑制效果；采用CPE法检测IFNα1b对登革2型（DENV-2）病毒的抑制效果、检测IFNα1b对EV71病毒的抑制效果、检测IFNα1b对人类腺病毒3型病毒株（AdV3）和人类腺病毒5型病毒株（AdV5）的抑制效果。采用空斑法检测IFNα1b对人类1型单纯疱疹病毒（HSV-1）的抑制效果。结果： IFNα1b对MDCK细胞的CC50值为479 572 U·mL-1，抑制H1N1的EC50为4 265 U·mL-1，选择指数（SI）为112.44，抑制H3N2的EC50为101 950 U·mL-1，SI为4.70，抑制B型流感病毒的EC50为18 240 U·mL-1，SI为18.50；IFNα1b对U937细胞的CC50值为大于100 000 U·mL-1，抑制H1N1病毒的EC50为37 897 U·mL-1，SI为大于2.64，抑制炎症因子IP-10的EC50为大于100 000 U·mL-1，抑制炎症因子IL-8的EC50为大于100 000 U·mL-1；IFNα1b和对LLC-MK2细胞的CC50值为大于10 000 U·mL-1，抑制HPIV3病毒的EC50为3.92 U·mL-1；IFNα1b对Hep 2细胞的CC50值为大于100 000 U·mL-1，抑制RSV病毒的EC50为1 661 U·mL-1，SI为大于60.2；IFNα1b对Vero E6细胞的CC50值为大于500 U·mL-1；IFNα1b对BHK-21细胞的CC50值为大于100 U·mL-1，抑制DENV-2病毒的EC50为0.90 U·mL-1，SI为大于111.11；IFNα1b对RD细胞的CC50值为大于100 000 U·mL-1，抑制EV71病毒的EC50为大于100 000 U·mL-1；IFNα1b对A549细胞的CC50值为大于100 000 U·mL-1，抑制AdV3病毒的EC50为100 000 U·mL-1，抑制AdV5病毒的EC50为100 000 U·mL-1；IFNα1b对Vero细胞的CC50值为大于100 000 U·mL-1，抑制HSV-1病毒的EC50为6 898 U·mL-1，SI为大于14.5。结论：在体外细胞水平上，IFNα1b 对A型流感病毒H1N1和H3N2、B型流感病毒、RSV病毒、DENV-2病毒、HPIV3病毒、HSV-1病毒均具有抑制作用，对EV71病毒、AdV3病毒和AdV5病毒无抑制作用，IFNα1b有望成为预防和治疗上呼吸道病毒感染的临床药物。

关键词: font-size:medium, ">人干扰素 α1b；体外；药效学；呼吸道病毒

Abstract: Objective： To evaluate the pharmacodynamic of human interferon α1b against broad-spectrum respiratory viruses in vitro. Methods： The toxicity of IFNα1b and positive control drugs on MDCK， U937， LLC-MK2， Hep-2， Vero E6， BHK-21， RD， A549 cells was detected by CPE method(cytopathic effect). Neuraminidase activity assay was used to detect the effects of IFNα1b and baloxavir on influenza virus strains CIV-H1N1/CNIC-1909(H1N1)， A/kansas/14/2017(NYMCX-327)(H3N2) and B/Maryland/15/2016(NYMCBX-69A) inhibitory effect； The inhibitory effects of IFNα1b， ribavirin and ibrutinib on influenza A strain(H1N1， A/PuertoRico/8/1934) and their inflammatory factors were detected by ELISA kit. Indirect immunofluorescence assay(IFA) was used to detect the inhibitory effect of IFNα1b and Ziresovir on respiratory syncytial virus(RSV A2) and the inhibitory effect of IFNα1b and ribavirin on human parainfluenza virus(HPIV3). The inhibitory effect of IFNα1b and NITD008 on DENV-2 virus， the inhibitory effect of IFNα1b and Vapendavir on EV71 virus， the inhibitory effect of IFNα1b and CMX001 on human adenovirus type 3 virus strain(AdV3) and human adenovirus type 5 were detected inhibitory effect of virus strain(AdV5). The inhibition effect of IFNα1b and acyclovir on human herpes simplex virus type 1(HSV-1) was measured by plaque assay.Results： The CC50 values of IFNα1b on MDCK cells were 479 572 U·mL-1. The EC50 values for inhibiting H1N1 were 4 265 U·mL-1. The selection index(SI) was 112.44. The EC50 for inhibiting H3N2 was 101 950 U·mL-1， and the SI was 4.70 The EC50 for inhibiting influenza B virus was 18 240 U·mL-1 and the SI was 18.50. The CC50 values of IFNα1b on U937 cells were more than 100 000 U·mL-1 and the EC50 values of H1N1 virus inhibition were 37 897 U·mL-1. The SI values were greater than 2.64 and the EC50 values for inhibiting inflammatory factor IP-10 were greater than 100 000 U·mL-1. The inhibitory EC50 values of IL-8 were more than 100 000 U·mL-1. The CC50 values of IFNα1b on LLC-MK2 cells were more than 10 000 U·mL-1 and the EC50 values of HPIV3 virus inhibition were 3.92 U·mL-1. The CC50 values of IFNα1b against Hep 2 cells were more than 100 000 U·mL-1. The EC50 values of RSV inhibition were 1661 U·mL-1 and the SI values were more than 60.2. The CC50 values of IFNα1b on BHK-21 cells were greater than 100 U·mL-1. The EC50 values of DENV-2 virus inhibition were 0.90 U·mL-1. The SI values were greater than 111.11. The CC50 values of IFNα1b on RD cells were greater than 100 000 U·mL-1. The EC50 values of EV71 inhibition were greater than 100 000 U·mL-1. The CC50 values of IFNα1b on A549 cells were greater than 100 000 U·mL-1 and the EC50 values of AdV3 virus inhibition were 100 000 U·mL-1. The EC50 was 100 000 U·mL-1. The CC50 values of IFNα1b for vero cell were more than 100 000 U·mL-1. The EC50 values for inhibiting HSV-1 virus were 6 898 U·mL-1 and the SI values were more than 14.5. Conclusion： At the cellular level in vitro， IFNα1b has inhibitory effects on influenza A viruses H1N1 and H3N2， influenza B viruses， RSV viruses， DENV-2 viruses， HPIV3 viruses and HSV-1 viruses. IFNα1b has no inhibitory effect on EV71 virus， AdV3 virus and AdV5 virus， and is expected to be a clinical drug for the prevention and treatment of upper respiratory tract viral infections.

Key words: font-size:medium, ">Human interferon α1b； Vitro； Pharmacodynamics； Respiratory viruses

中图分类号:

刘琳琳1, 王晓波1, 苏畅1, 张廷辉1, 刘玉林1, 刘景会1. 人干扰素α1b体外抗呼吸道病毒的药效学研究[J]. 中国药物评价, 2024, 41(3): 222-232.

LIU Lin-lin1, WANG Xiao-bo1, SU Chang1, ZHANG Ting-hui1, LIU Yu-lin1, LIU Jing-hui1. Human Interferon α1b Broad-spectrum Viruses Pharmacodynamics Study in Vitro[J]. CHINESE JOURNAL OF DRUG EVALUATION, 2024, 41(3): 222-232.

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