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

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。 尽管可获得FDA批准的佐剂，但对共佐剂的需要是明显的，因为单一佐剂 疫苗通常不产生持久的保护性免疫或免疫功能低下的免疫， 老年人口，这一点可以从每年发生的数千起死亡事件中得到证明， 流感疫苗。共佐剂是一种物质，其本身可能是或可能不是佐剂，但可以与联合收割机结合。 与已知的佐剂一起使用以提供协同效应，例如增强的抗体应答。利乐可 鉴定新的共佐剂集中在小分子上， 但是增强了由免疫抑制剂诱导的初级免疫激活，如核因子κ B（NF-κB）， TLR-4激动剂。我们开发了新的高通量筛选（HTS）方法，导致识别 当用Toll样受体-4激动剂LPS处理时，第一类延长NF-κB活化的化合物。的 用于HTS的表型测定产生了几种不同类型的化合物，其具有不同的代谢机制。 行动上一种这样的化学型四氢噻吩并[2，3-c]吡啶在我们的动力学中显示出最有效的活性 分析筛选和体内评价。因此，我们建议在这里探索构效关系 四氢噻吩并[2，3-c]吡啶类化合物的研究，目的是鉴定有效化合物 以及在支架上引入基团如芳基叠氮化物、生物素的位点，以获得亲和/光亲和探针。 然后将这些化合物在不同细胞中进行几种细胞因子诱导的体外测定， 评估作用机制并使用亲和/光亲和探针进行靶标鉴定。最后， 将在小鼠体内筛选有效化合物与FDA批准的佐剂单 磷酰脂质A（MPLA）使用模型抗原卵清蛋白来缩小到两种有效的化合物， 在鼠流感疫苗接种中进一步筛选，以观察用强毒剂量的 流感病毒、血凝素抑制和病毒中和滴度以及肺中的病毒滴度。

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.