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：
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
项目类别：
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

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.

肝炎病毒（HBV）的慢性感染是全球死亡的第七个主要原因，也是第五领先癌症的原因。 HBV慢性感染的关键是核局部HBV共价关闭驱动HBV转录和复制的圆形DNA（CCDNA）。 HCC的风险与 CCCDNA的复制水平。 HBV治疗需要消除CCCDNA，目前这是不可行的。但是，实现功能治疗，定义为乙型肝炎表面抗原（HBSAG）的持续丧失，可能通过沉默CCCDNA可能会有可能。当前的疗法，干扰素α或终身治疗维持核（T）IDE类似物的维护，功能性较低，强调了对新HBV的关键需求治疗。 HBV HBX蛋白由于其关键而成为抗病毒药的出色候选在病毒生命周期中的作用 - 病毒转录的调节，病毒限制因素的降解，该因子剪影因素 CCCDNA，干扰许多宿主细胞过程。尽管它具有核心作用，但HBX尚未我们的中心假设是靶向HBX将抑制病毒复制，轮廓 CCCDNA，并促进HBV功能固化。我们提出的研究基于最近的几项进展。第一的，我们已经显示HBX与细胞DDB1相互作用，以介导已知的静音蛋白的降解 CCCDNA。其次，我们使用人肝干细胞来开发了一个正宗的HBV感染模型器官（HLOS）。最后，我们与药物发现中心（CDD）建立了合作贝勒医学院已经开发了50个图书馆，其中包含超过50亿小说的DNA-bar编码小型药物样分子（DEC-TEC）。现在，我们提出了一个高度创新的项目，以最终确定新的HBV 靶向HBX的抗病毒药。在AIM 1中，纯化的HBX蛋白将通过相对于DEC的亲和力选择来筛选 TEC库。粘合剂将通过DNA测序鉴定，并用新鲜靶蛋白验证。考虑 HBX与100多种细胞蛋白相互作用，我们希望鉴定多个HBX粘合剂。结构活性关系（SAR）将使用计算软件确定，并测量粘合剂对HBX的亲和力。将追求具有SAR和高亲和力的化合物。在AIM 2中，将筛选高亲和力HBX粘合剂如在分裂荧光素酶测定中测量的临界HBX-DDB1相互作用的能力。复合然后，将测试抑制HBX-DDB1的HBX-DDB1，以抑制HLO复制模型中HBV复制的能力和与已知的核能（T）IDE类似物结构进行了比较。成功的HBX粘合剂将成为未来的体内评估，包括毒性，药物代谢和药代动力学。其他高亲和力粘合剂可以将靶向嵌合分子（protac）掺入蛋白酶体介导的降解。在全球范围内，超过2.57亿人患有HBV，大约有100万人感染每年肝肝硬化或癌症死亡。拟议的研究具有发现的巨大潜力治疗慢性HBV，预防HBV相关癌症并导致功能性治疗的新型方法。