Tetrahydrothieno[2,3-c]pyridines as Vaccine Co-adjuvants

四氢噻吩并[2,3-c]吡啶作为疫苗辅佐剂

• 批准号: 10307634
• 负责人: DENNIS CARSON
• 金额: $ 19.75万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-11-25 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10307634
• 关键词: Adjuvant; Adjuvant Study; Adjuvanticity; Affinity; Agonist; Antibody Response; Antibody titer measurement; Antigens; Azides; Binding; Biological Assay; Biological Markers; Biotin; Body Weight decreased; Bone Marrow; Bromides; Bronchoalveolar Lavage Fluid; California; Cell Line; Cells; Cessation of life; Dendritic Cells; Dose; Dyes; Elderly; FDA approved; Hemagglutinin; Human; Immune; Immunity; Immunization; Immunocompromised Host; In Vitro; Incubated; Influenza; Influenza Hemagglutinin; Influenza vaccination; Kinetics; Lead; Libraries; Ligands; Lipid A; Liposomes; Literature; Lung; Measures; Metabolic; Modeling; Modification; Mus; NF-kappa B; Nucleotides; Ovalbumin; Pattern recognition receptor; Phenotype; Poison; Population; Procedures; Proteins; Proteomics; Protocols documentation; Resources; Safety; Scheme; Site; Stimulus; Structure-Activity Relationship; TLR2 gene; TLR4 gene; Techniques; Toxic effect; Vaccine Adjuvant; Vaccines; Virulent; Virus; base; beta-Lactamase; cell type; cytokine; design; efficacy study; experimental study; high throughput screening; immune activation; in vitro Assay; in vivo; in vivo evaluation; influenza virus vaccine; influenzavirus; monocyte; mouse model; novel; pyridine; receptor; safety study; scaffold; screening; small molecule; ultraviolet irradiation

Project Summary Vaccines consisting of antigen and adjuvant rely primarily on adjuvants for enhancement of immune stimuli. Despite the availability of FDA approved adjuvants, the need for co-adjuvants is evident since single adjuvant vaccines often do not generate long lasting protective immunity or immunity in immunocompromised and elderly population as evidenced by the thousands of deaths that occur every year despite the availability of influenza vaccines. A co-adjuvant is a substance that may or may not be an adjuvant by itself but can combine with a known adjuvant to offer synergistic effects such as enhanced antibody response. Our approach towards identifying novel co-adjuvants has focused on small molecules that may not lead to immune activation by themselves but enhance the primary immune activation such as nuclear factor kappa B (NF-κB) induced by a TLR-4 agonist. We developed novel high-throughput screening (HTS) approaches that led to identification of first-in-class compounds that prolong NF-κB activation when treated with Toll-like receptor-4 agonist LPS. The phenotypic assay used for HTS yielded several different classes of compounds with different mechanisms of action. One such chemotype tetrahydrothieno[2,3-c]pyridines showed most potent activity in our kinetic profiling screening and in vivo evaluation. Thus, we propose here to explore structure-activity relationship studies in the tetrahydrothieno[2,3-c]pyridine class of compounds with an aim to identify potent compounds and sites on the scaffold to introduce groups such aryl azide, biotin to obtain affinity/photoaffinity probes. These compounds will then be subjected to several in vitro assays for cytokine inductions in different cells to evaluate mechanisms of action and perform target identification using the affinity/photoaffinity probes. Finally, the potent compounds will be screened in vivo in mice for co-adjuvanticity with FDA approved adjuvant mono phosphoryl lipid A (MPLA) using model antigen ovalbumin to narrow down to two potent compounds which will be further screened in murine influenza vaccination to observe for survival post challenge with virulent dose of influenza virus, hemagglutinin inhibition and virus neutralization titers as well as virus titers in the lung.

项目摘要 由抗原和佐剂组成的疫苗主要依靠佐剂来增强免疫刺激。 尽管FDA批准的佐剂已经上市，但由于单一佐剂，对辅助佐剂的需求是显而易见的 疫苗通常不能产生持久的保护性免疫或免疫受损和 老年人口--每年发生数千起死亡事件就是明证，尽管有 流感疫苗。辅助剂是一种物质，它本身可能是佐剂，也可能不是佐剂，但可以结合 用一种已知的佐剂来提供协同效应，例如增强抗体反应。我们的方法是 识别新的辅助剂专注于可能不会通过以下方式导致免疫激活的小分子 而增强核因子-κ-B等初级免疫活性。 TLR-4激动剂。我们开发了新的高通量筛选(HTS)方法，从而识别出 一流的化合物，在用Toll样受体-4激动剂脂多糖治疗时延长NF-κB的激活。这个 用于HTS的表型分析产生了几种不同类型的化合物，它们具有不同的作用机理 行动。一种这样的化学型四氢硫代并[2，3-c]吡啶在我们的动力学中显示出最强的活性 侧写筛选和体内评价。因此，我们建议在这里探索结构-活性关系 四氢硫代[2，3-c]吡啶类化合物的研究，旨在鉴定有效化合物 并在支架上引入芳基叠氮、生物素等基团，获得亲和/光亲和探针。 然后，这些化合物将在不同的细胞中接受几种细胞因子的体外诱导 评估作用机制并使用亲和/光亲和探针进行靶标识别。最后， 这些有效的化合物将在小鼠体内筛选与FDA批准的佐剂Mono的协同佐剂作用 使用模型抗原卵清蛋白缩小到两种有效化合物的磷酰脂A(MPLA)，这将 在小鼠流感疫苗接种中进一步筛查，观察毒力剂量攻击后的存活情况 流感病毒、血凝素抑制和病毒中和滴度以及肺中病毒滴度。