Discovery of inhibitors of EBV lytic cycle inducing protein ZTA for therapeutic development

发现 EBV 裂解周期诱导蛋白 ZTA 抑制剂用于治疗开发

基本信息

批准号：
10684643
负责人：
Takahiro Yano
金额：
$ 13.02万
依托单位：
VIRONIKA, LLC
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-16 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10684643
关键词：
Active Sites Address Advanced Development Affect Animal Model Autoimmune Diseases BZLF1 gene Binding Binding Proteins Biochemical Biological Assay Biophysics Biotechnology Burkitt Lymphoma Carcinogens Cells Characteristics Chronic Classification Collaborations Communicable Diseases Computer Models Computing Methodologies DNA DNA Binding DNA Binding Domain DNA-Binding Proteins Detection Development Disease Drug Kinetics EBV-associated disease Epstein-Barr Virus-Related Malignant Neoplasm Gene Expression Genes Goals Herpesviridae Hodgkin Disease Human Human Herpesvirus 4 Immune Immune System Diseases Immunocompromised Host Infectious Agent Infectious Mononucleosis Intercalating Agents Lead Libraries Life Cycle Stages Lymphomagenesis Lytic Phase Lytic Virus Malignant Neoplasms Measures Medical Membrane Proteins Methods Multiple Sclerosis Nasopharynx Carcinoma Natural Products Non-Hodgkin&apos s Lymphoma Nucleotides Oncogenic Oxidation-Reduction Pathology Periodicals Persons Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacodynamics Phase Population Production Proteins Quantitative Structure-Activity Relationship Research Research Project Grants Risk Factors Roentgen Rays Small Business Innovation Research Grant Stomach Carcinoma Structure Structure-Activity Relationship Surface Plasmon Resonance T-Cell Lymphoma Testing Therapeutic Agents Therapeutic Intervention Transcription Coactivator Universities Validation Viral Virus Diseases Z-Form DNA biophysical techniques carcinogenesis cell type design detection assay drug discovery experience follow-up high throughput screening improved inhibitor latent gene expression lead candidate lead series lytic gene expression nanomolar novel novel therapeutics plasma protein Z pre-clinical prevent programs screening skills small molecule small molecule inhibitor small molecule therapeutics targeted treatment therapeutic development tumorigenesis

项目摘要

Project Summary: The goal of this SBIR is to discover a small molecule inhibitor to the Epstein-Barr Virus (EBV)-encoded ZTA protein to prevent EBV-associated cancer and immune disorders. EBV is a human herpesvirus that switches between latent and lytic gene expression to escape immune detection and promote oncogenic transformation of diverse cell types. EBV is classified as a class I carcinogen by the WHO and estimated to be responsible for ~200,000 new cancer cases per year. EBV-associated cancers are diverse, and include subtypes of gastric carcinoma, nasopharyngeal carcinoma, Burkitt’s lymphoma, Hodgkin’s lymphoma, NK/T cell lymphoma, and various non-Hodgkin’s lymphomas particularly among immunocompromised populations. EBV is the major causative agent of infectious mononucleosis, and the predominant viral risk factor for multiple sclerosis (MS) and other auto immune diseases. The viral-encoded protein, ZTA (also known as BZLF1, ZEBRA, and Z) is a redox-sensitive DNA-binding transcriptional activator that is essential for EBV lytic cycle gene expression and viral production. The periodic cycling of lytic virus is a critical component for EBV-driven malignancy and auto- immune disease, and ZTA and the downstream viral and cellular genes that it activates, have been implicated in carcinogenesis and immune pathology. Therefore, ZTA is an attractive target for therapeutic intervention. The DNA-binding domain of ZTA has been characterized structurally and biochemically, and provides an ideal target for small molecule inhibition of EBV lytic life cycle. We have developed robust biochemical and cell-based assays for detection of ZTA DNA binding and used the biochemical assays for traditional high throughput screening (HTS). We screened the HTS hits using an unrelated DNA binding protein to eliminate DNA intercalators and tested the most selective compounds using surface plasmon resonance (SPR) to determine direct binding to the target protein. We now propose to use improved counter screening assays to characterize the hits from the previous screening efforts, expand our screening to natural products and an additional library of synthetic compounds designed for infectious disease targets. Natural products have advantageous characteristics such as higher rigidity for targeted protein surfaces outside of active sites like that of ZTA. We will use medicinal chemistry to define chemophore structure-activity relationships of our hits and identify a suitable lead compound for further development. Our goal is to produce lead compounds with nanomolar potency in biochemical assays, low micromolar activity in cell-based assays, and greater than 10-fold selectivity against control counter-screens. In Phase 2, we will develop our advanced hits into a pre-clinical lead candidate. The ultimate goal of this SBIR program is to develop a novel small molecule therapeutic agent to block the EBV lytic cycle and its associated diseases.

项目概要：该 SBIR 的目标是发现 Epstein-Barr 病毒 (EBV) 编码的 ZTA 的小分子抑制剂预防 EBV 相关癌症和免疫疾病的蛋白质 EBV 是一种可以转换的人类疱疹病毒。潜伏基因表达和裂解基因表达之间的关系，以逃避免疫检测并促进致癌转化 EBV 被世界卫生组织列为 I 类致癌物，估计是造成这种情况的原因。每年约 200,000 例新的 EBV 相关癌症病例多种多样，其中包括胃癌亚型。癌、鼻咽癌、伯基特淋巴瘤、霍奇金淋巴瘤、NK/T 细胞淋巴瘤和各种非霍奇金淋巴瘤，尤其是免疫功能低下人群中的 EBV 是主要的。传染性单核细胞增多症的病原体，以及多发性硬化症 (MS) 的主要病毒危险因素病毒编码的蛋白质 ZTA（也称为 BZLF1、ZEBRA 和 Z）是一种病毒编码蛋白。氧化还原敏感的 DNA 结合转录激活剂，对于 EBV 裂解周期基因表达至关重要裂解病毒的周期性循环是 EBV 驱动的恶性肿瘤和自身免疫性疾病的关键组成部分。免疫疾病、ZTA 及其激活的下游病毒和细胞基因都与此有关因此，ZTA 是治疗干预的一个有吸引力的靶点。 ZTA 的 DNA 结合域已在结构和生物化学上进行了表征，并提供了理想的 EBV 裂解生命周期的小分子抑制靶点。我们开发了强大的生化和细胞检测方法来检测 ZTA DNA 结合，并使用用于传统高通量筛选 (HTS) 的生化测定我们使用 HTS 筛选。不相关的 DNA 结合蛋白可消除 DNA 嵌入剂，并使用以下方法测试了最具选择性的化合物我们现在建议使用表面等离子共振（SPR）来确定与靶蛋白的直接结合。改进的反筛选测定来表征先前筛选工作的命中率，扩大我们的筛选天然产物和专为传染病设计的额外合成化合物库天然产物具有优势特性，例如目标蛋白质具有更高的刚性。我们将使用药物化学来定义化学团。我们的命中的结构-活性关系，并确定适合进一步开发的先导化合物。目标是生产在生化中具有纳摩尔效力、在分析中具有低微摩尔活性的先导化合物基于细胞的测定，以及比对照反筛选高出 10 倍的选择性。在第二阶段，我们将将我们的先进产品开发为临床前先导候选药物该 SBIR 计划的最终目标是开发一种新型小分子治疗剂来阻断 EBV 裂解周期及其相关疾病。