Antisense Mediated Treatment of Hepatitis B Virus Infection

乙型肝炎病毒感染的反义介导治疗

• 批准号: 8433308
• 负责人: Stefan Frank Wieland
• 金额: $ 72.22万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8433308
• 关键词: Accounting; Acute Hepatitis; Adverse effects; Animal Model; Antisense Technology; Antiviral Agents; Antiviral Therapy; Biological Models; Cell Culture Techniques; Characteristics; Chronic; Chronic Hepatitis; Chronic Hepatitis B; Clinical; Cytomegalovirus; DNA Viruses; Drug Kinetics; Drug resistance; Gene Expression; Goals; HBV Genotype; Hepatitis B Virus; Hepatitis C; Hepatocyte; Human; In Vitro; Infection; Institutes; Interferons; Kinetics; Laboratories; Lead; Life Cycle Stages; Liver; Liver Cirrhosis; Malignant neoplasm of liver; Mediating; Monitor; Mus; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Primary carcinoma of the liver cells; Production; Public Health; Rattus; Reagent; Regulation; Resistance; Serum; System; Testing; Therapeutic; Toxic effect; Transgenic Mice; Treatment Protocols; Vaccination; Vaccines; Viral; Viral Drug Resistance; Viral Genes; Viral Genome; Viremia; Virus Diseases; Virus Replication; Work; analog; base; combat; design; drug development; experience; fomivirsen; improved; in vivo; in vivo Model; interferon therapy; intrahepatic; killings; mouse model; mutant; novel; novel strategies; pathogen; pre-clinical; prevent; public health relevance; research study; response; socioeconomics; tool; transmission process; treatment planning; viral RNA

DESCRIPTION (provided by applicant): The hepatitis B virus (HBV) is a noncytopathic, hepatotropic DNA virus that causes acute and chronic hepatitis and kills a million people each year from cirrhosis of the liver and hepatocellular carcinoma. Currently approved therapies for the treatment of chronic hepatitis B include two classes of drugs: interferon and a small number of nucleos(t)ide analogues. However, the indication and response to interferon therapy is poor and associated with severe adverse effects and nucleos(t)ide analogues therapy is frequently associated with the emergence of single- and cross-drug-resistant HBV mutants. Thus, developing a new interferon- and/or nucleos(t)ide analogue-independent therapy is the long-term objective of this application. Antisense-mediated degradation of viral RNA represents such a novel class of antiviral therapy and this approach has been successfully pursued by our collaborator Isis Pharmaceuticals and resulted in a commercialized drug (fomivirsen) targeting cytomegalovirus. Using the same principal, our collaborators at Isis Pharmaceuticals have now identified lead candidate antisense molecules that efficiently inhibit HBV replication in cell culture. Studying the HBV life cycle and its inhibition by antiviral drugs has been and still is hampered by the lack of suitable and widely available in vivo HBV replication and infection systems. I have extensive experience using HBV transgenic mice that express and replicate HBV from an integrated viral genome to study viral gene expression, replication, virus production and inhibition of these steps by interferon-mediated antiviral mechanism. Furthermore, together with our collaborators at the Salk Institute, we have recently demonstrated efficient HBV and hepatitis C virus infection in a novel chimeric mouse model repopulated with human hepatocytes that allows us to study all steps of the viral life cycle during long-term infections. The goal of this application is to use these in vivo HBV replication and infection model systems to further evaluate and improve the therapeutic value and characteristics of the lead candidate antisense molecules targeting HBV. In specific aim 1 we will establish the lead candidate's toxicity and pharmacokinetic profile, demonstrate in vivo activity, and identify the most potent candidates. Specific aim 2 is designed to determine the long-term tolerability and efficacy of the lead candidates in suppressing HBV gene expression and replication in the HBV transgenic mice. In specific aim 3 the antiviral efficacy of the leads and the potential for emergence of drug resistance will be tested in the context of a bonefide HBV infection initiated with a monoclonal viral isolate using human hepatocyte repopulated chimeric mice. Finally, in specific aim 4 we will determine the leads antiviral efficacy against infection with patient sera containing isolates of different HBV genotypes and sera from patients that experienced virological breakthrough on antiviral therapy. Together, these studies will be instrumental in introducing antisense based therapy as a novel approach to combat chronic HBV infection.

描述(申请人提供)：乙肝病毒(乙肝病毒)是一种非细胞病变的嗜肝DNA病毒，可引起急性和慢性肝炎，每年导致100万人死于肝硬变和肝细胞癌。目前批准的治疗慢性乙肝的药物包括两类药物：干扰素和少量的核(T)类似物。然而，干扰素治疗的适应症和反应很差，并且与严重的不良反应相关，核苷(T)类似物治疗经常与单一和交叉耐药的乙肝病毒变异株的出现有关。因此，开发一种新的干扰素和/或核苷酸(T)类似物非依赖性治疗是该应用的长期目标。反义介导的病毒RNA降解代表了一种新型的抗病毒治疗，我们的合作者Isis PharmPharmticals已经成功地采用了这种方法，并导致了针对巨细胞病毒的商业化药物(福米韦森)。利用同样的原理，我们在Isis制药公司的合作者现在已经确定了有效抑制细胞培养中乙肝病毒复制的主要候选反义分子。由于缺乏合适和广泛可用的体内乙肝病毒复制和感染系统，研究乙肝病毒的生命周期及其抗病毒药物的抑制作用一直是并且仍然是阻碍的。我有丰富的经验，使用表达和复制乙肝病毒的转基因小鼠从完整的病毒基因组中研究病毒基因的表达、复制、病毒产生以及干扰素介导的抗病毒机制对这些步骤的抑制。此外，我们与索尔克研究所的合作伙伴最近在一种新的嵌合小鼠模型中展示了有效的乙肝和丙型肝炎病毒感染，该模型重新填充了人肝细胞，使我们能够研究长期感染期间病毒生命周期的所有步骤。这项应用的目的是利用这些体内乙肝病毒复制和感染模型系统来进一步评估和改进针对乙肝病毒的主要候选反义分子的治疗价值和特性。在具体目标1中，我们将建立主要候选药物的毒性和药代动力学特征，展示体内活性，并确定最有效的候选药物。特异靶2旨在确定候选药物在抑制乙肝转基因小鼠体内乙肝基因表达和复制方面的长期耐受性和有效性。在具体目标3中，将在使用人类肝细胞再繁殖嵌合小鼠的单克隆性病毒分离株引发的真正的乙肝病毒感染的背景下，测试这些先导的抗病毒效果和出现耐药性的可能性。最后，在特定的目标4中，我们将确定Leads的抗病毒效果，用含有不同HBV型分离株的患者血清和在抗病毒治疗方面取得病毒学突破的患者的血清来对抗感染。总之，这些研究将有助于将反义治疗作为对抗慢性乙肝感染的一种新方法。