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.
Public Health Relevance Statement: Project Narrative Epstein-Barr Virus (EBV) is responsible for ~200,000 new cancers and over 1 million new cases of infectious mononucleosis worldwide. EBV is the most prevalent viral risk factor for Multiple Sclerosis affecting over 400,000 persons per year in the US alone. EBV is classified as a human carcinogen associated with Burkitt's lymphoma, Hodgkin's lymphoma, nasopharyngeal carcinomas and gastric carcinomas. The product being developed in this proposal is the first therapeutic small molecule drug that specifically inhibits a key regulator of EBV life cycle.
Project Terms: Affect; inhibitor; Autoimmune Diseases; autoimmune condition; autoimmune disorder; Biological Assay; Assay; Bioassay; Biologic Assays; Biophysics; biophysical foundation; biophysical principles; biophysical sciences; Biotechnology; Biotech; Burkitt Lymphoma; Burkitt Tumor; Burkitt's Lymphoma/Leukemia; Burkitt's Type Small Non-Cleaved Cell Lymphoma; Malignant Neoplasms; Cancers; Malignant Tumor; malignancy; neoplasm/cancer; Carcinogens; Cancer Causing Agents; Oncogens; oncogenic agent; Cells; Cell Body; Pharmaceutical Chemistry; Medicinal Chemistry; Pharmaceutic Chemistry; Communicable Diseases; Infectious Disease Pathway; Infectious Diseases; Infectious Disorder; Computing Methodologies; computational methodology; computational methods; computer based method; computer methods; computing method; Crystallization; Disease; Disorder; DNA; Deoxyribonucleic Acid; DNA-Binding Proteins; Pharmaceutical Preparations; Drugs; Medication; Pharmaceutic Preparations; drug/agent; Human Herpesvirus 4; Burkitt Herpesvirus; Burkitt Lymphoma Virus; E-B Virus; EB virus; EBV; Epstein Barr Virus; HHV-4; HHV4; Infectious Mononucleosis Virus; Gene Expression; Genes; Goals; Herpesviridae; Herpesviruses; herpes virus; Hodgkin Disease; Hodgkin Disorder; Hodgkin lymphoma; Hodgkin's; Hodgkin's Lymphoma; Hodgkin's disease; Hodgkins lymphoma; Malignant Lymphogranuloma; Human; Modern Man; Immune System Diseases; Immune Diseases; Immune Disorders; Immune Dysfunction; Immune System Disorder; Immune System Dysfunction; Immune System and Related Disorders; Immunodeficiency and Immunosuppression Disorders; Immunologic Diseases; Immunological Diseases; Immunological Dysfunction; Immunological System Dysfunction; Infectious Mononucleosis; Glandular Fever; mononucleosis; Intercalating Agents; DNA Intercalating Agent; Intercalating Ligands; Intercalative Compounds; Intercalators; Lead; Pb element; heavy metal Pb; heavy metal lead; Libraries; Life Cycle Stages; Life Cycle; life course; Non-Hodgkin's Lymphoma; Nonhodgkins Lymphoma; non-Hodgkins disease; Membrane Proteins; Membrane Protein Gene; Membrane-Associated Proteins; Surface Proteins; Methods; Multiple Sclerosis; Disseminated Sclerosis; insular sclerosis; Persons; Nucleotides; Redox; oxidation reduction reaction; Oxidation-Reduction; Pathology; Cyclicity; Rhythmicity; Periodicity; Pharmacokinetics; Drug Kinetics; Production; Proteins; Research; Risk Factors; Plant Embryos; Plant Zygotes; seed; Seeds; chemical structure function; structure function relationship; Structure-Activity Relationship; Testing; Universities; Virus Diseases; Viral Diseases; viral infection; virus infection; virus-induced disease; Roentgen Rays; X-Radiation; X-Ray Radiation; X-ray; Xray; T-Cell Lymphoma; T-Cell Non-Hodgkin's Lymphoma; T-Cell NonHodgkins Lymphoma; T-Cell and NK-Cell Non-Hodgkin's Lymphoma; Measures; Immunocompromised Host; Immunocompromised; Immunocompromised Patient; Immunosuppressed Host; immunosuppressed patient; base; improved; Chronic; Phase; Biochemical; Medical; Active Sites; Nasopharyngeal Carcinoma; Nasopharynx Carcinoma; Ligand Binding Protein; Ligand Binding Protein Gene; Protein Binding; bound protein; Binding Proteins; R-Series Research Projects; R01 Mechanism; R01 Program; Research Grants; Research Projects; Research Project Grants; Collaborations; Therapeutic Agents; infectious organism; Infectious Agent; programs; Immunes; Immune; cell type; Viral; Cancer Induction; carcinogenesis; experience; Surface Plasmon Resonance; Animal Models and Related Studies; model of animal; model organism; Animal Model; Structure; skills; novel; Gastric Carcinoma; Stomach Carcinoma; intervention therapy; Therapeutic Intervention; Pharmacodynamics; High Throughput Assay; high throughput screening; QSAR; Quantitiative Structure Activity Relationship; Quantitative Structure-Activity Relationship; drug discovery; virus related cancer; Virus-Related Malignancy; Virus-Related Malignant Neoplasm; viral associated cancer; viral associated malignancy; viral associated malignant neoplasm; viral induced cancer; viral induced malignancy; viral induced malignant neoplasm; viral related cancer; viral related malignancy; viral related malignant neoplasm; virus associated cancer; virus associated malignancy; virus associated malignant neoplasm; virus induced cancer; virus induced malignancy; virus induced malignant neoplasm; DNA Binding Interaction; DNA bound; DNA Binding; Molecular Interaction; Binding; preventing; prevent; Oncogenesis; tumorigenesis; small molecule; Transcription Activator; Transcription Factor Coactivator; Transcriptional Activator; Transcriptional Activator/Coactivator; Transcriptional Coactivator; transcription co-activator; transcriptional co-activator; Transcription Coactivator; Address; DNA-Binding Protein Motifs; DNA Binding Domain; Detection; Lytic Phase; Lytic Cycle; Lytic Infection; Lytic Virus; Small Business Innovation Research Grant; SBIR; Small Business Innovation Research; Validation; Characteristics; follow-up; Active Follow-up; active followup; follow up; followed up; followup; Development; developmental; pre-clinical; preclinical; Advanced Development; design; designing; BZLF1 gene; BZLF1; BZLF1 protein; Epstein-Barr virus BZLF1 protein; Epstein-Barr virus Zta protein; ZEBRA protein; Zta protein; Zta transcription activator; lytic gene expression; EBV-associated disease; Epstein-Barr Virus associated disease; Lymphomagenesis; Population; Oncogenic; novel therapeutics; new drug treatments; new drugs; new therapeutics; new therapy; next generation therapeutics; novel drug treatments; novel drugs; novel therapy; therapeutic development; therapeutic agent development; lead series; screening; biophysical techniques; biophysical approaches; biophysical methodology; biophysical methods; targeted treatment; targeted drug therapy; targeted drug treatments; targeted therapeutic; targeted therapeutic agents; targeted therapy; Natural Products; small molecule inhibitor; small molecule therapeutics; nanomolar; nano-molar; lead candidate; Computer Models; Computerized Models; computational modeling; computational models; computer based models; computerized modeling; detection assay
