肝细胞亲环的Alisporivir抑制HBV减少复制和乙肝表面抗原生产

StephenW

Alisporivir Inhibition of Hepatocyte Cyclophilins Reduces HBV Replication and Hepatitis B Surface Antigen Production
Sandra Phillips∗
,
Shilpa Chokshi∗
,
Udayan Chatterji
,
Antonio Riva
,
Michael Bobardt
,
Roger Williams
,
Philippe Gallay
,
Nikolai V. Naoumov correspondence email
∗Authors share co-first authorship.
Received: December 15, 2013; Accepted: October 5, 2014; Published Online: October 08, 2014
DOI: http://dx.doi.org/10.1053/j.gastro.2014.10.004


Background & Aims

Cyclophilins are host factors required for hepatitis C virus replication. Cyclophilin inhibitors such as alisporivir have shown strong anti–hepatitis C virus activity in vitro and in clinical studies. However, little is known about whether hepatocyte cyclophilins are involved in the hepatitis B virus (HBV) life cycle. We investigated the effects of 2 cyclophilin inhibitors (alisporivir and NIM811) on HBV replication and hepatitis B surface antigen (HBsAg) production in cell lines.
Methods

Liver-derived cell lines producing full-length HBV and HBsAg particles, owing to stable (HepG2215) or transient (HuH-7) transfection, or infected with HBV (HepaRG cells; Invitrogen [Carlsbad, CA]), were incubated with alisporivir or NIM811 alone, or alisporivir in combination with a direct antiviral (telbivudine). The roles of individual cyclophilins in drug response was evaluated by small interfering RNA knockdown of cyclophilin (CYP)A, CYPC, or CYPD in HepG2215 cells, or CYPA knockdown in HuH-7 cells. The kinetics of antiviral activity were assessed based on levels of HBV DNA and HBsAg and Southern blot analysis.
Results

In HepG2215, HuH-7, and HepaRG cells, alisporivir reduced intracellular and secreted HBV DNA, in a dose-dependent manner. Knockdown of CYPA, CYPC, or CYPD (reduced by 80%) significantly reduced levels of HBV DNA and secreted HBsAg. Knockdown of CYPA significantly reduced secretion of HBsAg, leading to accumulation of intracellular HBsAg; the addition of alisporivir greatly reduced levels of HBsAg in these cells. The combination of alisporivir and telbivudine had greater antiviral effects than those of telbivudine or alisporivir alone.
Conclusions

Alisporivir inhibition of cyclophilins in hepatocyte cell lines reduces replication of HBV DNA and HBsAg production and secretion. These effects are potentiated in combination with direct antiviral agents that target HBV-DNA polymerase.
Keywords:
Cyclophilin Inhibition, Direct-Acting Antiviral Agent, Peptidyl-Prolyl Isomerase, Viral Replication

StephenW

肝细胞亲环的Alisporivir抑制HBV减少复制和乙肝表面抗原生产
桑德拉·菲利普斯*
，
希尔帕Chokshi*
，
查特吉主席Udayan
，
李安东
，
迈克尔·Bobardt
，
罗杰·威廉姆斯
，
菲利普Gallay
，
尼古拉五Naoumov对应的电子邮件
*作者份额共同第一作者。
收稿日期：二〇一三年十二月十五日;接受：2014年10月5日;网上公布：2014年10月8日
DOI：http://dx.doi.org/10.1053/j.gastro.2014.10.004


背景与目的

亲环所需的丙型肝炎病毒复制的主机因素。亲环蛋白抑制剂如alisporivir已经在体外和临床研究中显示了强大的抗丙型肝炎病毒活性。然而，鲜为人知的是，肝细胞亲环是否涉及乙型肝炎病毒（HBV）的生命周期。我们研究了2亲环蛋白抑制剂（alisporivir和NIM811）对HBV复制和B型肝炎表面抗原（HBsAg）的生产细胞系的影响。
方法：

生产全长HBV和HBsAg颗粒，由于稳定（HepG2215）或瞬变（的HuH-7）的转染或感染HBV肝脏来源的细胞系（HepaRG细胞; Invitrogen公司[卡尔斯巴德，CA）]，孵育alisporivir或NIM811独自一人，或alisporivir与直接抗病毒（替比夫定）的组合。在药物反应个体亲环的作用是由亲环素的小干扰RNA击倒评价（CYP）A，长江电力，或CYPD在HepG2215细胞或CYPA击倒在HuH-7细胞。抗病毒活性的动力学基于HBV DNA和HBsAg和Southern印迹分析的水平进行评估。
结果

在HepG2215，的HuH-7和HepaRG细胞，alisporivir降低细胞内和细胞分泌的HBV DNA，以剂量依赖的方式。 CYPA，长江电力，或CYPD（减少80％）的击倒显著降低的HBV DNA的水平和分泌的HBsAg。 CYPA击倒显著降低的HBsAg分泌，导致细胞内的HBsAg的积累;加入alisporivir大大降低的HBsAg水平在这些细胞中。 alisporivir和替比夫定的结合有比夫定或alisporivir单独的更大的抗病毒效果。
结论

在肝细胞系Alisporivir抑制亲环的降低的HBV DNA和HBsAg的产生和分泌的复制。这些效果增效与靶向的HBV-DNA聚合酶直接抗病毒剂的组合。
关键词：
亲环素抑制，直接作用抗病毒药剂，肽基脯氨酰异构酶，病毒复制

StephenW

旧的研究，刚刚出版
OnCore的乙肝候选（OCB-030）被设计为抑制亲环
OCB-030的定进入临床试验在今年下半年

战天斗hbv

感谢分享，展望2015

newchinabok

NVP018, A CYCLOPHILIN INHIBITOR FOR TREATMENT OF CHRONIC HBV INFECTION       

J. Nilsson1, S. Moss2, N. Coates2, M. Bobardt3, P. Gallay3, M. Gregory2
1Neurovive Pharmaceutical AB, Lund, Sweden, 2Isomerase Therapeutics Ltd., Cambridge, United Kingdom, 3The Scripps Research Institute, La Jolla, CA, United States
E-mail: matt.***@****

Background and aims: Whilst cyclophilin (Cyp) is a clinically validated target for chronic HCV infection, recent work has shown that inhibition of Cyps by either siRNA knockdown or therapeutically can also inhibit hepatitis B virus in vitro. NVP018 (previously known as BC556) is an orally available sangamide-based cyclophilin inhibitor (CI), with a well-differentiated preclinical profile when compared to cyclosporine-based CIs such as alisporivir and SCY-635. It has a very different structure and does not have the liabilities associated with these cyclosporine-based compounds.
Methods: Analysis of NVP018 viral inhibition, including mechanism of action studies and immune modulation in models of hepatitis B virus infection.
Results: NVP018 is a potent inhibitor of HBV replication distinct from previously studied inhibitors. It has a dual mechanism of action including both direct inhibition of viral pathways which require cyclophilins and immune modulation via IRFs, including potent inhibition of an interaction between cyclophilin A and IRF9, a key component of the Jak/Stat pathway.
Conclusions: NVP018 inhibits HBV infection in various models via new mechanisms and represents a novel combination partner for nucleotide inhibitors or interferons for treatment of chronic HBV infection, with the potential to inhibit viral replication and increase seroconversion rates.

newchinabok

NVP018，A环素抑制剂治疗慢性乙肝病毒感染

J. Nilsson1，S. Moss2，N. Coates2，M. Bobardt3，P. Gallay3，M. Gregory2
1Neurovive制药AB，瑞典隆德，2Isomerase治疗有限公司，剑桥，英国，3The斯克里普斯研究所，拉霍亚，CA，美国
电子邮件：matt.***@****

背景和目的：尽管环素（CYP）是一种临床验证的目标为慢性丙型肝炎病毒感染，最近的工作已表明，抑制的CYP的由任一siRNA敲除或治疗还可以抑制乙肝病毒在体外。 NVP018（以前称为BC556）是一种口服可用sangamide基环素抑制剂（CI）的，具有分化良好的临床前轮廓相比，环孢素基证明书如alisporivir和SCY-635。它具有非常不同的结构，并且不与这些环孢菌素类化合物有关的债务。
方法：NVP018病毒抑制，包括行动研究中的乙型肝炎病毒感染的模型机理和免疫调制分析。
结果：NVP018是HBV复制的有效抑制剂从先前研究的抑制剂明显。它具有双重作用机制包括直接抑制需要亲环和经由脉冲响应函数免疫调节，包括亲环素A和IRF9的JAK / STAT途径的一个关键组成部分之间的相互作用的强效抑制病毒途径。
结论：NVP018抑制HBV感染，通过新的机制，各种型号和代表核苷酸酶抑制剂或干扰素一个新的合作伙伴联合治疗慢性HBV感染，具有抑制病毒复制，增加血清转换率的潜力。

战天斗hbv

一直不理解亲环素抑制剂的优势在哪，望解释

newchinabok

靶点是人体，不是病毒，病毒要入侵人体，要和人体细胞发生关系，阻断人体细胞某些必须因子，也叫宿主抑制剂，不是病毒抑制剂，也就间接抑制病毒

战天斗hbv

回复 newchinabok 的帖子

哦。。。。这个也是HIV那边延伸过来的技术吧

StephenW

A New Approach to Hepatitis B Therapy?
165 Liver/Biliary 2 days ago Kristine Novak 1

Cyclophilins are involved in multiple steps of the hepatitis B virus (HBV) life cycle in hepatocytes—cyclophilin inhibitors reduce viral replication and HBV envelope protein production and secretion, researchers report in the February issue of Gastroenterology. The cyclophilin inhibitor alisporivir, combined with the HBV polymerase inhibitor, reduces markers of HBV infection and HBV replication in cells, revealing a possible new therapeutic approach for chronic hepatitis B.
Hepatitis B virions. Source: CDC



Cyclophilins have peptidyl-prolyl isomerase activity, and catalyze the cis to trans conversion of proline-containing peptides and facilitate protein folding. There are several human genes that encode cyclophilins, including CYPA, CYPB, CYPC, and CYPD. Their products localize to different areas of the cell. For example, CYPA and CYP40 are present in the cytosol, CYPB and CYPC in the lumen of the endoplasmic reticulum, and CYPD is found in the mitochondria.

Cyclophilins regulate the life cycle of a range of viruses including HCV, HIV, vaccinia virus, coronaviruses, and polyomavirus BK, and are required for virus replication. CYPA is the main cyclophilin that regulates the life cycle of HCV; cyclophilins reduce HCV replication and secretion from infected cells.

Sandra Phillips et al investigated whether hepatocyte cyclophilins are involved in HBV replication, hepatitis B surface antigen (HBsAg) production, and secretion in liver-derived cell lines that produce full-length HBV and HBsAg particles.

They found that the cyclophilin inhibitor alisporivir reduced intracellular and secreted HBV DNA in liver cells lines in a dose-dependent manner. Knockdown of CYPA, CYPC, or CYPD significantly reduced levels of HBV DNA and secreted HBsAg—a marker of HBV infection.

Small molecule inhibitors of cyclophilins also reduced levels of HBsAg produced by cells, and caused a dose-dependent reduction in HBV DNA. The combination of alisporivir and telbivudine had greater antiviral effects than those of telbivudine or alisporivir alone. The authors propose that the cyclophilin inhibitors alisporivir and NIM811 block multiple cyclophilins, affecting not only HBV replication and HBsAg production, but multiple stages of the HBV life cycle.

CYPA is one of the most abundant cytosolic proteins (approximately 0.1% of cell proteins). Phillips et al showed that CYPA participates in HBV replication and envelope protein secretion from hepatocytes. They say that CYPA is important for the formation of intracellular, nucleocapsid-associated HBV DNA and viral secretion and for HBsAg secretion from cells.

The authors propose a new approach for treating chronic hepatitis B that involves a combination of a direct antiviral agent (such as the HBV-DNA polymerase inhibitors) and agents that act directly on liver cells, such as those that block cyclophilins.

Alisporivir is in development for the treatment of chronic hepatitis C. Phillips et al say that it has been evaluated in more than 2000 patients and seems to be well tolerated, with a better safety profile when given as interferon-free treatment than with interferon.

The authors conclude that combination treatment with an HBV nucleos(t)ide polymerase inhibitor plus a cyclophilin inhibitor would interfere with multiple sites of the HBV life cycle, reducing HBsAg production and secretion as well as blocking HBV entry into non-infected hepatocytes.