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: 8243502
• 负责人: Stefan Frank Wieland
• 金额: $ 76.83万
• 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/8243502
• 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. PUBLIC HEALTH RELEVANCE: Over 350 million people throughout the world are chronically infected by HBV, 15-25% of whom will die from cirrhosis of the liver and liver cancer. The work described in this proposal will exploit current and novel in vivo model systems to discover improved, safe and effective antiviral drugs to alleviate the human suffering and socioeconomic burden of this threat to public health.

描述（由申请人提供）：B肝炎病毒（HBV）是一种非致细胞病变的嗜肝DNA病毒，可引起急性和慢性肝炎，每年有100万人死于肝硬化和肝细胞癌。目前批准用于治疗慢性B型肝炎的疗法包括两类药物：干扰素和少量核苷（酸）类似物。然而，干扰素治疗的适应症和反应较差，并伴有严重的不良反应，核苷（酸）类似物治疗经常与单药和交叉耐药HBV突变体的出现相关。因此，开发新的干扰素和/或核苷（酸）类似物非依赖性疗法是本申请的长期目标。反义介导的病毒RNA降解代表了这样一类新型的抗病毒治疗，我们的合作者Isis Pharmaceuticals已经成功地采用了这种方法，并产生了一种靶向巨细胞病毒的商业化药物（福米韦生）。使用相同的原理，我们在Isis Pharmaceuticals的合作者现在已经确定了在细胞培养中有效抑制HBV复制的主要候选反义分子。研究HBV的生命周期和抗病毒药物的抑制作用一直受到缺乏合适的和广泛可用的体内HBV复制和感染系统的阻碍。我有丰富的经验，使用HBV转基因小鼠表达和复制HBV从一个完整的病毒基因组研究病毒基因表达，复制，病毒生产和抑制这些步骤的干扰素介导的抗病毒机制。此外，我们与Salk研究所的合作者一起，最近在一种新的嵌合小鼠模型中证明了有效的HBV和丙型肝炎病毒感染，该模型重新填充了人类肝细胞，使我们能够研究长期感染期间病毒生命周期的所有步骤。本申请的目的是使用这些体内HBV复制和感染模型系统来进一步评估和改善靶向HBV的先导候选反义分子的治疗价值和特征。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，证明体内活性，并确定最有效的候选药物。具体目标2旨在确定主要候选药物在HBV转基因小鼠中抑制HBV基因表达和复制的长期耐受性和疗效。在具体目标3中，将使用人肝细胞再增殖嵌合小鼠，在单克隆病毒分离株引发的真正HBV感染的背景下，检测先导化合物的抗病毒疗效和出现耐药性的可能性。最后，在具体目标4中，我们将确定先导化合物对抗含有不同HBV基因型分离株的患者血清和来自抗病毒治疗中经历病毒学突破的患者血清感染的抗病毒疗效。总之，这些研究将有助于引入基于反义的治疗作为对抗慢性HBV感染的新方法。 公共卫生相关性：全世界有超过3.5亿人慢性感染HBV，其中15-25%将死于肝硬化和肝癌。本提案中描述的工作将利用当前和新型的体内模型系统来发现改进的、安全的和有效的抗病毒药物，以减轻这种对公共卫生的威胁所带来的人类痛苦和社会经济负担。