抑制HepaRG细胞和原代人肝细胞中核酸聚合物的乙型肝炎病毒

StephenW

Inhibition of hepatitis B viral entry by nucleic acid polymers in HepaRG cells and primary human hepatocytes

    Clément Guillot,
    Nora Martel,
    Françoise Berby,
    Isabelle Bordes,
    Olivier Hantz,
    Matthieu Blanchet,
    Camille Sureau,
    Andrew Vaillant ,
    Isabelle Chemin

Inhibition of hepatitis B viral entry by nucleic acid polymers in HepaRG cells and primary human hepatocytes

   
PLOS x

    Published: June 21, 2017
    https://doi.org/10.1371/journal.pone.0179697


Abstract

Hepatitis B virus (HBV) infection remains a major public health concern worldwide with 240 million individuals chronically infected and at risk of developing cirrhosis and hepatocellular carcinoma. Current treatments rarely cure chronic hepatitis B infection, highlighting the need for new anti-HBV drugs. Nucleic acid polymers (NAPs) are phosphorothioated oligonucleotides that have demonstrated a great potential to inhibit infection with several viruses. In chronically infected human patients, NAPs administration lead to a decline of blood HBsAg and HBV DNA and to HBsAg seroconversion, the expected signs of functional cure. NAPs have also been shown to prevent infection of duck hepatocytes with the Avihepadnavirus duck hepatitis B virus (DHBV) and to exert an antiviral activity against established DHBV infection in vitro and in vivo.

In this study, we investigated the specific anti-HBV antiviral activity of NAPs in the HepaRG human hepatoma cell line and primary cultures of human hepatocytes. NAPs with different chemical features (phosphorothioation, 2’O-methyl ribose, 5-methylcytidine) were assessed for antiviral activity when provided at the time of HBV inoculation or post-inoculation. NAPs dose-dependently inhibited HBV entry in a phosphorothioation-dependent, sequence-independent and size-dependent manner. This inhibition of HBV entry by NAPs was impaired by 2’O-methyl ribose modification. NAP treatment after viral inoculation did not elicit any antiviral activity.

StephenW

抑制HepaRG细胞和原代人肝细胞中核酸聚合物的乙型肝炎病毒进入

    ClémentGuillot，
    诺拉·马特尔，
    FrançoiseBerby，
    伊莎贝尔·博尔德（Isabelle Bordes）
    Olivier Hantz，
    Matthieu Blanchet，
    卡米尔局，
    安德鲁·威兰特，
    Isabelle Chemin

抑制HepaRG细胞和原代人肝细胞中核酸聚合物的乙型肝炎病毒进入

   
PLOS x

    发布时间：2017年6月21日
    https://doi.org/10.1371/journal.pone.0179697


抽象

乙型肝炎病毒（HBV）感染仍然是全球主要的公共卫生问题，其中2.4亿人长期感染并发生肝硬化和肝细胞癌的风险。目前的治疗很少治愈慢性乙型肝炎感染，强调需要新的抗HBV药物。核酸聚合物（NAP）是硫代磷酸化的寡核苷酸，其已经证明具有抑制几种病毒感染的巨大潜力。在长期感染的人类患者中，NAPs导致血液HBsAg和HBV DNA的下降以及HBsAg血清学转换，预期功能治愈的迹象。 NAP也被证明可以防止禽流感病毒鸭乙型肝炎病毒（DHBV）感染鸭肝细胞，并在体外和体内对已建立的DHBV感染发挥抗病毒活性。

在本研究中，我们调查了HepaRG人肝癌细胞系和人肝细胞原代培养物中NAP的特异性抗HBV抗病毒活性。在HBV接种或接种后提供时，评估具有不同化学特征（硫代磷酸化，2'O-甲基核糖，5-甲基胞苷）的NAP的抗病毒活性。 NAP以硫代磷酸依赖性，与序列无关和大小依赖的方式剂量依赖地抑制HBV进入。 NAPs对HBV进入的抑制受到2'O-甲基核糖修饰的损害。病毒接种后的NAP治疗未引起任何抗病毒活性。

StephenW

Replicor announces publication of its studies on the antiviral activity of NAPs in recognized in vitro models of HBV infection



MONTREAL, June 28, 2017 – Replicor Inc., a privately held biopharmaceutical company targeting a cure for patients with chronic hepatitis B virus (HBV) and chronic HBV and hepatitis delta virus (HDV) co-infection, announced today the publication of its study on the activity of NAPs in recognized in vitro models of HBV infection in the journal PLOS ONE. (http://journals.plos.org/plosone ... &type=printable).

This study assessed the antiviral activity of a variety of NAP compounds, including the clinically active NAPs REP 2055, REP 2139 and REP 2165, in HBV infected HepaRG and primary human hepatocytes in tissue culture.  These studies demonstrated that early generation NAPs like REP 2055 had an ability to block viral entry only at high concentrations (5-10μM) while later generation (RNA) NAPs like REP 2139 and REP 2165 had negligible entry effect.  Dr. Andrew Vaillant, CSO of Replicor commented, “This study is a classic example of the known challenges of working with this chemical class in tissue culture”, adding “the post-entry effects of NAPs we know to be present could not be observed in these models because the intracellular trafficking of phosphorothioate oligonucleotides (NAPs are in this class) that occurs in patients is absent in tissue culture.”

The study did provide some important advances, demonstrating that the antiviral effects of REP 2139 and REP 2165 in the currently ongoing REP 401 trial are not driven by entry inhibition. Dr. Vaillant went on to state, “This latest study tells us we are focusing our research efforts in the right direction in developing a tissue culture model of HBV infection where the intracellular NAP transit occurring in hepatocytes in patients is restored.  The initial results of these efforts were presented at EASL 2017 (http://replicor.com/wp-content/u ... SL-2017-THU-156.pdf) and we will continue to refine our ability to replicate the post-entry effects of NAPs in vitro.”

About Replicor

Replicor is a privately held biopharmaceutical company with the most advanced animal and human clinical data in the development of the cure for HBV and HDV. The company is dedicated to accelerating the development of an effective treatment for patients with HBV and HBV/HDV co-infection. For further information about Replicor please visit our website at www.replicor.com.

StephenW

复制者宣布其公认的HBV感染体外模型中NAP的抗病毒活性研究



蒙特利尔，2017年6月28日 - Replicor Inc.是一家私人控股的生物制药公司，旨在治愈慢性乙型肝炎病毒（HBV）和慢性乙型肝炎病毒（HBV）和乙型肝炎病毒（HDV）共感染患者，今天宣布发表研究报告关于国家行动方案在公认的体外HBV感染模型中的活动。 （http://journals.plos.org/plosone ... &type=printable）。

本研究评估了各种NAP化合物的抗病毒活性，包括在组织培养中HBV感染的HepaRG和原代人肝细胞中临床活跃的NAP REP2055，REP2139和REP2165。这些研究表明早期的NAP如REP 2055具有仅在高浓度（5-10μM）下阻断病毒进入的能力，而后代（RNA）NAP如REP 2139和REP 2165具有可忽略的进入效应。 Replicor CSO博士Andrew Vaillant博士评论说：“这项研究是组织培养中化学类工作中已知挑战的典型例子”，并补充说：“我们知道存在的NAP的进入后效应是无法观察的在这些模型中，因为在组织培养中不存在发生在患者中的硫代磷酸寡核苷酸（NAPs在该类别中）的细胞内运输。

该研究确实提供了一些重要进展，表明REP 2139和REP 2165在目前正在进行的REP 401试验中的抗病毒作用不受入侵抑制的驱动。 Vaillant博士接着说：“这项最新的研究告诉我们，我们正在把我们的研究工作集中在正确的方向，以开发HBV感染的组织培养模型，其中患者肝细胞中发生的细胞内NAP转运恢复。这些努力的初步结果在EASL 2017上提供（http://replicor.com/wp-content/u ... SL-2017-THU-156.pdf）我们将继续提高我们在体外复制NAPs后进入效应的能力。“

关于复制者

复制者是一家私人持有的生物制药公司，拥有最先进的动物和人类临床资料，用于治疗HBV和HDV。该公司致力于加速HBV​​和HBV / HDV共感染患者的有效治疗发展。有关Replicor的更多信息，请访问我们的网站www.replicor.com。