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

基本信息

批准号：
10451632
负责人：
Sue Ellen Crawford
金额：
$ 20万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-15 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10451632
关键词：
Affinity Animal Model Antiviral Agents Antiviral Therapy Apoptosis Area Bar Codes Binding 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 Core Antigen Hepatitis B Infection 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 Persons Pharmaceutical Preparations Pre-Clinical Model Primary carcinoma of the liver cells Production Protac Proteins 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 antiviral drug development 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

项目摘要

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.

乙型肝炎病毒 (HBV) 慢性感染是全球第七大死亡原因，也是第五大死亡原因 HBV慢性感染的关键是核定位的HBV附加型共价闭合。驱动 HBV 转录和复制的环状 DNA (cccDNA) 与 HCC 风险直接相关。 cccDNA 的复制水平乙肝病毒的治愈需要消除 cccDNA，但目前这是不可行的。然而，实现功能性治愈（定义为乙型肝炎表面抗原 (HBsAg) 持续丧失）可能会目前的疗法是通过干扰素α或终生治疗来沉默cccDNA。维持核苷（酸）类似物的功能性治愈率较低，凸显了对新型乙型肝炎病毒的迫切需求 HBV HBx 蛋白因其关键性而成为开发抗病毒药物的绝佳候选者。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尽管 HBx 具有核心作用，但它尚未成为最重要的因素。我们的中心假设是，针对 HBx 会抑制病毒复制、沉默。 cccDNA，并促进 HBV 功能性治愈。首先，我们提出的研究建立在几项最新进展的基础上。我们已经证明 HBx 与细胞 DDB1 相互作用，介导已知沉默的细胞蛋白的降解其次，我们利用人肝干细胞开发了真实的 HBV 感染模型。最后，我们与药物发现中心 (CDD) 建立了合作。贝勒医学院开发了 50 个库，包含超过 50 亿条新型 DNA 条形码小分子我们现在提出一个高度创新的项目，以最终识别新的 HBV。目标 HBx 的抗病毒药物在目标 1 中，将通过针对 DEC- 的亲和力选择来筛选纯化的 HBx 蛋白。 Tec 文库将通过 DNA 测序进行鉴定，并使用新鲜的目标蛋白进行验证。由于 HBx 与 100 多种细胞蛋白相互作用，我们期望鉴定出多种 HBx 结合物的结构-活性。关系（SAR）将使用计算软件确定，并测量结合剂对HBx的亲和力。在目标 2 中，将寻找具有 SAR 和高亲和力的化合物，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-DDB1 相互作用的能力。然后将在 HLO 复制模型中测试抑制 HBx-DDB1 抑制 HBV 复制的能力，与已知的核苷（酸）类似物恩替卡韦相比，成功的 HBx 结合剂将作为先导药物。未来的体内评估，包括毒性、药物代谢和药代动力学。可以掺入蛋白酶体介导的靶向嵌合分子 (PROTAC) 的蛋白水解作用全世界有超过 2.57 亿人慢性感染 HBV，其中大约有 100 万人。每年都有人死于肝硬化或癌症。拟议的研究对于发现肝硬化或癌症具有巨大的潜力。治疗慢性乙型肝炎、预防乙型肝炎相关癌症并实现功能性治愈的新方法。