Small molecule inhibitors of HBx that decrease hepatitis B virus replication

减少乙型肝炎病毒复制的 HBx 小分子抑制剂

• 批准号: 10284389
• 负责人: Sue Ellen Crawford
• 金额: $ 24万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10284389
• 关键词: Affinity; Animal Model; Antiviral Agents; Antiviral Therapy; Apoptosis; Area; Bar Codes; Binding; Biological; Biological Assay; Biological Process; Cancer Etiology; Categories; Cause of Death; Cell Cycle Progression; Cell Nucleus; Cell physiology; Chromosomal Stability; Chronic; Chronic Hepatitis B; Circular DNA; Cirrhosis; Clinical; Clinical Trials; Collaborations; Computer software; DNA; DNA Sequence; DNA sequencing; Development; Drug Kinetics; Drug Targeting; Drug usage; Epitopes; Future; Genetic Transcription; Goals; HBV Genotype; Hepatitis B; Hepatitis B Core Antigen; Hepatitis B Surface Antigens; Hepatitis B Virus; Hepatocyte; Human; Infection; Interferon-alpha; Knowledge; Lead; Libraries; Life; Life Cycle Stages; Liver Cirrhosis; Liver Stem Cell; Luciferases; Maintenance; Malignant Neoplasms; Malignant neoplasm of liver; Measures; Mediating; Medicine; Messenger RNA; Methods; Modeling; Natural Immunity; Nuclear; Organoids; Pathway interactions; Patients; Pharmaceutical Preparations; Pre-Clinical Model; Primary carcinoma of the liver cells; Production; Proteins; Proteolysis; Regulation; Reverse Transcriptase Inhibitors; Risk; Role; Signal Transduction; Structure-Activity Relationship; Technology; Testing; Therapeutic; Time; Toxic effect; Viral; Viral Proteins; Virus; Virus Diseases; Virus Replication; analog; base; chronic infection; clinically relevant; college; cost; drug discovery; drug metabolism; drug resistant virus; entecavir; first-in-human; genetic regulatory protein; in vivo evaluation; innovation; multicatalytic endopeptidase complex; multidisciplinary; mutant; novel; novel strategies; novel therapeutics; pre-clinical; prevent; protein degradation; recruit; side effect; small molecule; small molecule inhibitor; virus related cancer

Chronic infection with hepatitis B virus (HBV) is the 7th leading cause of death worldwide and the 5th leading cause of cancer. The key to HBV chronic infection is the nuclear localized HBV episomal covalently closed circular DNA (cccDNA) that drives HBV transcription and replication. The risk of HCC is directly related to the level of replication from the cccDNA. An HBV cure requires eliminating cccDNA, which is not feasible at present. However, achieving a functional cure, defined as a sustained loss of hepatitis B surface antigen (HBsAg), may be possible by silencing the cccDNA. The current therapies, treatment with interferon alpha or life-long maintenance on nucleos(t)ide analogs, have low functional cure rates highlighting the critical need for new HBV therapeutics. The HBV HBx protein is an excellent candidate for the development of antivirals due to its critical roles in the virus life cycle – regulation of viral transcription, degradation of viral restriction factors that silence cccDNA, and interference with many host cellular processes. Despite its central role, HBx has yet to be the target of antiviral therapy. Our central hypothesis is that targeting HBx will inhibit viral replication, silence cccDNA, and facilitate an HBV functional cure. Our proposed studies build on several recent advances. First, we have shown HBx interacts with cellular DDB1 to mediate the degradation of cellular proteins known to silence cccDNA. Second, we have developed an authentic HBV infection model using human liver stem cell-derived organoids (HLOs). Finally, we have established a collaboration with the Center for Drug Discovery (CDD) at Baylor College of Medicine that has developed 50 libraries containing over 5 billion novel DNA-bar coded small drug-like molecules (DEC-Tec). We now propose a highly innovative project to ultimately identify new HBV antivirals that target HBx. In Aim 1, purified HBx protein will be screened by affinity selection against the DEC- Tec libraries. Binders will be identified by DNA sequencing and validated with fresh target protein. Considering that HBx interacts with over 100 cellular proteins, we expect to identify multiple HBx binders. Structure-activity relationship (SAR) will be determined using computational software, and the binder's affinity for HBx measured. Compounds with SAR and high affinity will be pursued. In Aim 2, high affinity HBx binders will be screened for the ability to inhibit the critical HBx-DDB1 interaction as measured in a split luciferase assay. Compounds that inhibit HBx-DDB1 will then be tested for the ability to inhibit HBV replication in the HLO replication model and compared against the known nucleos(t)ide analogue entecavir. Successful HBx binders will serve as leads for future in vivo evaluation that includes toxicity, drug metabolism, and pharmacokinetics. Other high affinity binders can be incorporated into proteolysis targeting chimeric molecules (PROTACs) for proteasome-mediated degradation. Worldwide, over 257 million people are chronically infected with HBV and approximately 1 million die each year of liver cirrhosis or cancer. The proposed studies hold tremendous potential for the discovery of novel approaches to treat chronic HBV, prevent HBV-associated cancer, and lead to a functional cure.

慢性B型肝炎病毒（HBV）感染是全球第7大死亡原因，也是全球第5大死亡原因。 癌症的原因。HBV慢性感染的关键是核定位的HBV游离体共价闭合 环状DNA（cccDNA）驱动HBV转录和复制。HCC的风险与 cccDNA的复制水平。HBV治疗需要消除cccDNA，这在目前是不可行的。 然而，达到功能性治愈，定义为持续丧失B型肝炎表面抗原（HBsAg），可能 通过沉默cccDNA是可能的。目前的治疗方法，用干扰素α或终身治疗 维持核苷（酸）类似物，功能性治愈率低，突出了对新HBV的迫切需求 治疗学HBV HBx蛋白是开发抗病毒药物的极好候选者，因为其关键的生物学特性。 在病毒生命周期中的作用-调节病毒转录，降解沉默的病毒限制因子 cccDNA，并干扰许多宿主细胞过程。尽管HBx发挥着核心作用，但它尚未成为 抗病毒治疗靶点。我们的中心假设是，靶向HBx将抑制病毒复制，沉默 cccDNA，并促进HBV功能性治愈。我们提出的研究建立在几个最近的进展。第一、 我们已经证明HBx与细胞DDB 1相互作用，介导已知沉默的细胞蛋白质的降解， cccDNA。第二，我们已经开发了一个真实的HBV感染模型，使用人肝干细胞衍生的 类器官（HLO）。最后，我们与药物发现中心（CDD）建立了合作关系， 贝勒医学院已经开发了50个图书馆，其中包含超过50亿个新的DNA条形码小 药物样分子（DEC-Tec）。我们现在提出一个高度创新的项目，以最终确定新的HBV 针对HBx的抗病毒药物在目的1中，将通过针对DEC-1的亲和选择来筛选纯化的HBx蛋白。 技术图书馆。将通过DNA测序鉴定结合剂，并使用新鲜靶蛋白进行验证。考虑 HBx与超过100种细胞蛋白相互作用，我们期望鉴定多种HBx结合剂。构效 将使用计算软件确定结合剂与HBx的亲和力的关系（SAR），并测量结合剂对HBx的亲和力。 将追求具有SAR和高亲和力的化合物。在目标2中，将筛选高亲和力HBx结合剂， 在裂解荧光素酶测定中测量的抑制关键HBx-DDB 1相互作用的能力。的化合物 然后在HLO复制模型中测试抑制HBx-DDB 1抑制HBV复制的能力， 与已知的核苷类似物恩替卡韦相比。成功的HBx粘合剂将作为以下方面的线索 未来的体内评价，包括毒性、药物代谢和药代动力学。其他高亲和力结合剂 可整合到蛋白酶体介导的蛋白酶解靶向嵌合分子（PROTAC）中 降解在世界范围内，超过2.57亿人慢性感染HBV，约100万人感染HBV。 每年死于肝硬化或癌症。拟议的研究对发现 治疗慢性HBV、预防HBV相关癌症并导致功能性治愈的新方法。