疫苗接种可以帮助预防感染，还可以用于治疗癌症、过敏，甚至可能治疗药物过量。佐剂可以增强疫苗反应，但目前，其进步和发展的道路是渐进的。我们使用 THP-1 细胞进行表型小分子筛选来鉴定核因子-κB (NF-κB) 激活分子，然后使用原代人外周血单核细胞通过定量肿瘤坏死因子免疫分析对先导靶标库进行反筛选。对原代细胞的筛选鉴定出一种咪唑并嘧啶，称为 PVP-037。此外，虽然 PVP-037 没有明显激活 THP-1 细胞，但它表现出广泛的先天免疫激活作用，包括体外原代人白细胞的 NF-κB 和细胞因子诱导，以及流感和 SARS-CoV-2 抗原特异性的增强小鼠的体液反应。多项从头合成结构增强迭代提高了 PVP-037 的体外功效、效力、物种特异性活性和体内佐剂性。总体而言，我们发现咪唑并嘧啶 Toll 样受体 7/8 佐剂与水包油乳剂协同作用，可增强免疫反应。

Vaccination can help prevent infection and can also be used to treat cancer, allergy, and potentially even drug overdose. Adjuvants enhance vaccine responses, but currently, the path to their advancement and development is incremental. We used a phenotypic small-molecule screen using THP-1 cells to identify nuclear factor-κB (NF-κB)-activating molecules followed by counterscreening lead target libraries with a quantitative tumor necrosis factor immunoassay using primary human peripheral blood mononuclear cells. Screening on primary cells identified an imidazopyrimidine, dubbed PVP-037. Moreover, while PVP-037 did not overtly activate THP-1 cells, it demonstrated broad innate immune activation, including NF-κB and cytokine induction from primary human leukocytes in vitro as well as enhancement of influenza and SARS-CoV-2 antigen-specific humoral responses in mice. Several de novo synthesis structural enhancements iteratively improved PVP-037's in vitro efficacy, potency, species-specific activity, and in vivo adjuvanticity. Overall, we identified imidazopyrimidine Toll-like receptor-7/8 adjuvants that act in synergy with oil-in-water emulsion to enhance immune responses.