Screening and identification of compounds with antiviral activity against hepatitis B virus using a safe compound library and novel real-time immune-absorbance PCR-based high throughput system.
Screening and identification of compounds with antiviral activity against hepatitis B virus using a safe compound library and novel real-time immune-absorbance PCR-based high throughput system.
复制标题
DOI：
10.1016/j.antiviral.2013.02.001
复制
发表时间：
2013-04
影响因子：
7.6
通讯作者：
Lu, Xuanyong
中科院分区：
文献类型：
--
作者：
Lamontagne, Jason;Mills, Courtney;Mao, Richeng;Goddard, Cally;Cai, Dawei;Guo, Haitao;Cuconati, Andy;Block, Timothy;Lu, Xuanyong
关键词：
现在有7种核苷/核苷类似物，沿着干扰素-α，被FDA批准用于治疗慢性肝炎B病毒（HBV）感染，这种疾病影响全世界数亿人。然而，这些药物的有效性有限，并且显著的副作用和病毒逃逸突变体的出现使得开发新的和更新的治疗方法成为治疗HBV的迫切需要。考虑到这一点，测试了一个含有2，000种已知在人类和小鼠中安全的化合物的文库，这些化合物在哺乳动物细胞中具有已知的作用机制，其可能具有抗HBV的抗病毒活性或在HBV产生细胞系中具有选择性毒性。一种改良的实时免疫吸收聚合酶链反应（IA-PCR）检测开发了这个屏幕，利用细胞，生产和分泌完整的HBV病毒粒子。在此过程中，病毒颗粒首先被固定在平板上的抗-HBs抗体捕获。随后通过实时PCR直接在捕获的颗粒上评估病毒载量。使用该测定，在对细胞活力没有可检测影响的条件下，8种化合物显示出一致地减少培养基中分泌的HBV病毒颗粒的量。两种化合物，丙美卡因和叶绿酸，被证明可以降低HBV水平4- 6倍，IC 50分别为1和1.5μM，并被选择用于进一步研究。这些化合物作为有希望的抗HBV的抗病毒药物候选物的鉴定，尽管缺乏先前对HBV抗病毒活性的认识，支持了测试已知化合物针对HBV的“病原体外靶标”活性的有效性和实用性，并且还验证了该IA-PCR测定作为检测针对包膜病毒的抗病毒活性的重要工具。
There are now 7 nucleoside/tide analogues, along with interferon-α, that are approved by the FDA for the management of chronic hepatitis B virus (HBV) infection, a disease affecting hundreds of millions of people worldwide. These medications, however, are limited in usefulness, and significant side effects and the emergence of viral escape mutants make the development of novel and updated therapeutics a pressing need in the treatment of HBV. With this in mind, a library containing 2,000 compounds already known to be safe in both humans and mice with known mechanisms of action in mammalian cells were tested for the possibility of either antiviral activity against HBV or selective toxicity in HBV producing cell lines. A modified real-time immune-absorbance-polymerase chain reaction (IA-PCR) assay was developed for this screen, utilizing cells that produce and secrete intact HBV virions. In this procedure, viral particles are first captured by an anti-HBs antibody immobilized on a plate. The viral load is subsequently assessed by real-time PCR directly on captured particles. Using this assay, eight compounds were shown to consistently reduce the amount of secreted HBV viral particles in the culture medium under conditions that had no detectable impact on cell viability. Two compounds, proparacaine and chlorophyllide, were shown to reduce HBV levels 4- to 6-fold with an IC50 of 1 and 1.5μM respectively, and were selected for further study. The identification of these compounds as promising antiviral drug candidates against HBV, despite a lack of previous recognition of HBV antiviral activity, supports the validity and utility of testing known compounds for “off- pathogen target” activity against HBV, and also validates this IA-PCR assay as an important tool for the detection of anti-viral activity against enveloped viruses.
登录
查看更多内容
影响因子：
4.9
作者：
Ladner, SK;Otto, MJ;King, RW
通讯作者：
King, RW
DOI：
10.1073/pnas.0912966107
发表时间：
2010-01-05
影响因子：
11.1
作者：
Gastaminza, Pablo;Whitten-Bauer, Christina;Chisari, Francis V.
通讯作者：
Chisari, Francis V.
影响因子：
24.5
作者：
JANSSEN, HLA;BERK, L;MULDER, CJJ
通讯作者：
MULDER, CJJ
影响因子：
3.8
作者：
Kartal, Elif Doyuk;Nayman, Saygin;Usluer, Gaye
通讯作者：
Usluer, Gaye
DOI：
10.1016/s1499-3872(12)60144-1
发表时间：
2012-04-01
影响因子：
3.3
作者：
Wu, Feng;Wu, Min-Jin;Zhu, Biao
通讯作者：
Zhu, Biao