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

描述（由申请方提供）：长期目标是探索一类具有独特作用机制的新型化合物用于治疗B型肝炎（HBV）的潜力，该作用机制不同于现有的抗病毒药物。HBV是感染肝脏并导致严重后果的传染性因子，例如肝衰竭、纤维化、肝硬化和肝细胞癌。所有批准的化疗治疗（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体总是随着长期治疗而出现。因此，需要具有不同于对病毒DNA聚合酶的作用的机制的新化合物，其单独起作用或与当前治疗组合起作用。我们从天然产物向日葵黄素中合成并发现了一系列具有广谱抗病毒活性的衍生物。两个衍生物，8-1和5-4-2，在培养中表现出有效的抗HBV活性。与目前临床上使用的仅抑制HBV DNA合成的抗HBV药物不同，8-1和5-4-2抑制HBV DNA、RNA和蛋白质，如在HBV稳定转染的细胞系中测定的。对拉米夫定耐药的HBV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HBV药物，未见报道。作用机制似乎涉及直接改变病毒转录复合物。在本申请中，提出了如下两个具体目的：1. 8-1和5-4-2抗HBV作用的独特机制研究。这包括：抗HBV RNA转录的机制、增强抗HBV活性的核苷类药物、抗HBeAg- HBV的活性以及HBV感染细胞的摄取和代谢。2.评价8-1的体内抗HBV活性。5-4-2号营地将作为备用营地一种新的HBV转基因小鼠将用于获得8-1的初步药代动力学信息和毒性，以及评估8-1的抗病毒活性。所提出的研究不仅将提供关于新化合物8-1作为抗HBV药物的潜力的信息，而且还将提供关于肝脏富集的转录因子在调节HBV转录中的作用的信息。

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.