A Novel Inhibitor of Human La Protein with Anti-HBV Activity Discovered by Struc

StephenW

A Novel Inhibitor of Human La Protein with Anti-HBV Activity Discovered by Structure-Based Virtual Screening and In Vitro Evaluation

Jing Tang1, Zhi-Min Huang2, Ying-Yi Chen2, Zhao-Hui Zhang3, Gao-Lin Liu1*, Jian Zhang2*

1 Department of Pharmacy, First People's Hospital Affiliated with Shanghai JiaoTong University, Shanghai, China, 2 Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis, Chinese Ministry of Education, Shanghai JiaoTong University School of Medicine, Shanghai, China, 3 Department of Urology, Ruijin Hospital Luwan Branch Affiliated with Shanghai JiaoTong University School of Medicine, Shanghai, China

Abstract
Background
Over 350 million people worldwide are infected with hepatitis B virus (HBV), a major cause of liver failure and hepatocellular carcinoma. Current therapeutic agents are highly effective, but are also associated with development of viral resistance. Therefore, strategies for identifying other anti-HBV agents with specific, but distinctive mechanisms of action are needed. The human La (hLa) protein, which forms a stabilizing complex with HBV RNA ribonucleoprotein to promote HBV replication, is a promising target of molecular therapy.
Aims
This study aimed to discover novel inhibitors of hLa that could inhibit HBV replication and expression.
Methods
A multistage molecular docking approach was used to screen a Specs database and an in-house library against hLa binding sites. Sequential in vitro evaluations were performed to detect potential compounds with high scores in HepG2.2.15 cells.
Results
Of the 26 potential compounds with high scores chosen for experimental verification, 12 had HBV DNA inhibition ratios of less than 50% with P<0.05. Six had significant inhibition of HBV e antigen (HBeAg) levels, and 13 had significant inhibition of HBV surface antigen (HBsAg) levels by in vitro assays. Compounds HBSC-11, HBSC-15 and HBSC-34 (HBSC is system prefix for active compounds screened by the library) were selected for evaluation. HBSC-11 was found to have an obvious inhibitory effect on hLa transcription and expression.
Conclusions
Our findings suggest that anti-HBV activity of HBSC-11 may be mediated by a reduction in hLa levels. In addition, our data suggest the potential clinical use of hLa inhibitors, such as HBSC-11, for treating HBV infection.

StephenW

背景

全球超过350万人感染了乙型肝炎病毒（HBV），肝功能衰竭和肝癌的主要病因。目前的治疗药物是非常有效的，但也与病毒抗性的发展。因此，确定具体的行动，但独特的机制与其他抗乙肝病毒药物的战略是必要的。人La蛋白（HLA），形成稳定与HBV RNA的核糖促进HBV复制的复杂，是一个有前途的目标分子疗法。
旨在

本研究旨在发现HLA小说的抑制剂，能抑制HBV复制和表达。
方法

采用多级分子对接方法筛选规格数据库和对人类白细胞抗原结合位点的房子库。在体外评估顺序进行检测HepG2.2.15细胞中的高分潜力的化合物。
结果

实验验证选择高分的26个潜在的化合物中，有12例HBV DNA的抑制率低于50％，以P<0.05。 6例乙肝病毒e抗原（HBeAg）水平显着抑制，13例乙肝病毒表面抗原（HBsAg）水平显着的抑制作用，在体外实验。化合物，货源货源-11-15-34货源（货源是系统的活性化合物筛选库的前缀）被选定为评估。货源-11被发现有明显的抑制作用HLA转录和表达。
结论

我们的研究结果表明，抗乙肝病毒活性货源-11可减少HLA各级调解。此外，我们的数据表明HLA抑制剂潜在的临床使用，如货源-11，用于治疗乙肝病毒感染。

StephenW

HBSC11 seems to be able to inhibit HbsAg, HbeAg and hbvdna replication.
HBSC11似乎能够抑制HBsAg，HBeAg和HBVDNA复制
Of the 26 selected compounds, only HBSC-11, HBSC-15 and HBSC-34 were found to significantly decrease HBeAg, HBsAg and HBV DNA levels (Figures 3A–C). Therefore, these three compounds were selected to further evaluate their inhibition of hLa levels in HepG2.2.15 cells. Finally, only one compound, HBSC-11, was shown to have significant inhibitory effects on hLa expression, as evidenced by Western blotting (Figure 3D, HBSC-11 vs Untreated cells = 20.42% vs 100%, P = 0.0069; siRNA vs Untreated cells = 28.59% vs 100%, P = 0.0302) and qRT-PCR (Figure 3E, HBSC-11 vs Untreated cells = 29.73% vs 100%, P = 0.0217; siRNA vs Untreated cells = 19.42% vs 100%, P = 0.0116).
最后，只有一个化合物，HBSC-11，HLA表达上有显着的抑制作用，Western印迹证明（图3D，HBSC-11比未经处理的细胞=20.42％和100％，P =0.0069;的siRNA比未处理细胞=28.59％和100％，P =0.0302）和实时定量PCR（图3E，HBSC-11比未经处理的细胞=29.73％和100％，P =0.0217;的siRNA比未经处理的细胞=19.42％和100％，磷= 0.0116）。

StephenW

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