药学学报, 2019, 54(9): 1582-1587
引用本文:
陈秀秀, 罗荣华, 郑昌博, 姚债文, 唐秋菊, 熊思东, 郑永唐. 富马酸替诺福韦二吡呋酯体外抗寨卡病毒活性研究[J]. 药学学报, 2019, 54(9): 1582-1587.
CHEN Xiu-xiu, LUO Rong-hua, ZHENG Chang-bo, YAO Zhai-wen, TANG Qiu-ju, XIONG Si-dong, ZHENG Yong-tang. The anti-ZIKA virus activity of tenofovir disoproxil fumarate in vitro[J]. Acta Pharmaceutica Sinica, 2019, 54(9): 1582-1587.

富马酸替诺福韦二吡呋酯体外抗寨卡病毒活性研究
陈秀秀1,2,4, 罗荣华2, 郑昌博3, 姚债文1,2,4, 唐秋菊3, 熊思东4, 郑永唐2,4
1. 苏州大学药学院, 江苏 苏州 215021;
2. 中国科学院昆明动物研究所 云南省活性多肽研究与利用重点实验室, 云南 昆明 650223;
3. 昆明医科大学药学院暨云南省天然药物药理重点实验室, 云南 昆明 650500;
4. 苏州大学-中国科学院昆明动物研究所疾病动物模型与新药研发联合实验室, 江苏 苏州 215021
摘要:
富马酸替诺福韦二吡呋酯(tenofovir disoproxil fumarate,TDF)是一种核苷类似物(nucleoside analogues),已广泛用于临床治疗人类免疫缺陷病毒(human immunodeficiency virus,HIV)和乙型肝炎病毒(hepatitis B virus,HBV)感染。本研究旨在探讨TDF是否具有体外抗寨卡病毒(Zika virus,ZIKV)活性。首先通过噬斑抑制实验在细胞水平检测TDF是否具有抑制ZIKV作用,随后通过实时荧光定量PCR(real-time quantitative PCR)实验和蛋白质印迹(Western blot)实验分别在RNA水平和蛋白水平进一步验证TDF的抗ZIKV活性,最后通过MTT实验检测TDF的细胞毒性。结果发现,TDF不仅可降低ZIKV感染细胞后噬斑形成,还可抑制ZIKV RNA复制和ZIKV NS2B蛋白的表达,其对ZIKV的半数有效浓度(50% effective concentration,EC50)为14.96~27.47μmol·L-1。阳性药物利巴韦林抑制ZIKV的EC50为56.01 ±12.16 μmol·L-1。MTT检测结果表明,TDF和利巴韦林的细胞毒性很小,半数细胞毒浓度(50% cytotoxic concentration,CC50)均大于500 μmol·L-1。通过CC50/EC50计算出TDF的治疗指数(therapeutic index,TI)大于18.20,高于阳性对照药利巴韦林。本研究表明,TDF在细胞、RNA及蛋白水平均具有很好的体外抗ZIKV活性,有望成为抗ZIKV治疗的候选药物。
关键词:    富马酸替诺福韦二吡呋酯      核苷类似物      寨卡病毒      抗病毒活性      治疗     
The anti-ZIKA virus activity of tenofovir disoproxil fumarate in vitro
CHEN Xiu-xiu1,2,4, LUO Rong-hua2, ZHENG Chang-bo3, YAO Zhai-wen1,2,4, TANG Qiu-ju3, XIONG Si-dong4, ZHENG Yong-tang2,4
1. College of Pharmacy, Soochow University, Suzhou 215021, China;
2. Key Laboratory of Bioactive Peptides of Yunnan Province, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650223, China;
3. School of Pharmaceutical Science and Yunnan Key Laboratory of Pharmacology for Natural Products, Kunming Medical University, Kunming 650500, China;
4. KIZ-SU Joint Laboratory of Animal Model and Drug Development, College of Pharmaceutical Sciences, Soochow University, Suzhou 215021, China
Abstract:
Tenofovir disoproxil fumarate (TDF) is a nucleoside analogue that has been widely used for clinical treatment of human immunodeficiency virus (HIV) and hepatitis B virus (HBV) infection. The aim of this study was to investigate whether TDF has anti-Zika virus (ZIKV) activity in vitro. The inhibitory effect of TDF on ZIKV was detected by plaque reduction assay. Then, the anti-ZIKV activity of TDF at RNA level and protein level was verified by real time quantitative PCR and Western blot. Finally, MTT assay was used to determine the cytotoxicity of TDF. Our results showed that TDF not only reduced the formation of plaque after ZIKV infection, but also inhibited the replication of ZIKV RNA or expression of ZIKV NS2B protein. The 50% effective concentration (EC50) of TDF in inhibition of ZIKV replication were 14.96-27.47 μmol·L-1, while that of ribavirin was 56.01 ±12.16 μmol·L-1, which served as the positive control. The cytotoxicity of TDF and ribavirin in Vero cells were very low, with their 50% cytotoxic concentration (CC50) values being greater than 500 μmol·L-1. The therapeutic index of TDF calculated by CC50/EC50 was greater than 18.20, which was significantly higher than that of ribavirin. The results suggest that TDF has good anti-ZIKV activity in vitro and is expected to become a candidate drug for anti-ZIKV therapy.
Key words:    tenofovir disoproxil fumarate    nucleoside analogue    Zika virus    antiviral activity    therapy   
收稿日期: 2018-12-23
DOI: 10.16438/j.0513-4870.2018-1135
基金项目: 国家重点研发计划课题(2016YFC1201000);中国科学院重点部署项目课题(ZDRW-ZS-2016-4-2);云南省活性多肽研究与利用重点实验室开放课题(AMHD-2017-3).
通讯作者: 郑永唐,Tel/Fax:86-871-5195684,E-mail:zhengyt@mail.kiz.ac.cn
Email: zhengyt@mail.kiz.ac.cn
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