Mechanism and in vivo anti-HBV study of a novel class of non-nucleoside compounds

描述(申请人提供)：长期目标是探索一类新型化合物的潜力，该化合物具有与现有抗病毒药物不同的独特作用机制，用于治疗乙肝(乙肝)。乙肝病毒是一种感染性物质，会感染肝脏并导致严重后果，如肝功能衰竭、纤维化、肝硬变和肝细胞癌。所有已获批准的化疗药物(即拉米夫定、阿德福韦和恩替卡韦)仅通过干扰病毒DNA合成来抑制病毒复制。由于病毒DNA聚合酶突变而产生的耐药变异总是随着长期的治疗而出现。因此，除了对病毒DNA聚合酶的作用外，需要具有其他作用机制的新化合物，无论是单独作用还是与当前的治疗方法结合使用。我们合成了一系列天然产物辣黄素的衍生物，并发现它们具有广谱的抗病毒活性。8-1和5-4-2两个衍生物在培养条件下表现出较强的抗乙肝病毒活性。与目前临床使用的仅抑制HBVDNA合成的抗HBV药不同，8-1和5-4-2可抑制HBVDNA、RNA和蛋白质，如在稳定表达HBVDNA的细胞系中检测到的那样。对拉米夫定耐药的乙肝病毒对这些化合物仍然敏感。8-1和5-4-2的作用不同于其他抗乙肝药物，未见报道。其作用机制似乎涉及病毒转录复合体的直接改变。在这项应用中，提出了两个具体的目的：1.研究8-1和5-4-2抗病毒作用的独特机制。这包括：抗HBVRNA转录的机制，抗HBV核苷类药物的增强作用，抗HBeAg-HBV病毒的活性，以及乙肝病毒感染细胞的摄取和代谢。2.评价8-1的体内抗乙肝病毒活性。化合物5-4-2将作为后备化合物。8-1的药代动力学信息和毒性以及评价8-1的抗病毒活性将被用于一种新的乙肝转基因小鼠。这些研究不仅将提供新化合物8-1作为抗乙肝药物的潜力，还将提供肝脏富集型转录因子在调节乙肝病毒转录中的作用。

DESCRIPTION (provided by applicant): The long term goal is to explore the potential of a novel class of compounds with a unique mechanism of action different from existing antiviral drugs for the treatment of hepatitis B (HBV). HBV is an infectious agent that infects the liver and causes severe consequences, such as liver failure, fibrosis, cirrhosis and hepatocellular carcinoma. All the approved chemotherapeutic treatments (i.e. Lamivudine, Adefovir and Entecavir) only inhibit viral replication by interfering viral DNA synthesis. Resistant variants which develop due to viral DNA polymerase mutation invariably emerge with long-term treatment. Therefore, novel compounds with mechanisms other than the action on viral DNA polymerase, working alone or in combination with current treatments, are needed. A series of derivatives from the natural product, Helioxanthin, were synthesized and discovered to perform a wide-spectrum of antiviral activity by us. Two derivatives, 8-1 and 5-4-2, exhibited potent anti-HBV activity in culture. Unlike the current clinically used anti-HBV drugs that only inhibited HBV DNA synthesis, 8-1 and 5-4-2 inhibit HBV DNA, RNA, and proteins as determined in HBV stably transfected cell lines. HBV resistant to Lamivudine was still sensitive to these compounds. The action of 8-1 and 5-4-2 is different from other anti-HBV drugs and has never been reported. The mechanism of action seems to involve direct alteration of the viral transcription complex. In this application two specific aims are proposed as following: 1. Study of the unique mechanism of the antiviral action of 8-1 and 5-4-2 against HBV. This includes: the mechanism against HBV RNA transcription, potentiation of anti-HBV activity nucleoside drugs, activity against HbeAg- HBV, and uptake and metabolism in HBV infected cells. 2. Evaluation of the in vivo anti-HBV activity of 8-1. Compound 5-4-2 will be the backup compound. A novel HBV transgenic mice will be used to obtain preliminary pharmacokinetic information and toxicity of 8-1 as well as to assess antiviral activity of 8-1. The studies proposed will provide information about not only the potential of the novel compound 8-1, as an anti-HBV drug, but also the roles of liver enriched transcription factors in regulation of HBV transcription.