Paradiso Emiliano, Janež Špela, Jakopin Žiga
Faculty of Pharmacy, Department of Pharmaceutical Chemistry, University of Ljubljana, Aškerčeva 7, SI-1000 Ljubljana, Slovenia.
ACS Omega. 2025 Aug 22;10(34):39060-39072. doi: 10.1021/acsomega.5c05358. eCollection 2025 Sep 2.
Novel immunopotentiators are essential for advancing our understanding of immune receptor crosstalk and for addressing infectious diseases. Previous studies have suggested that coactivation of nucleotide-binding oligomerization domain-containing protein 2 (NOD2) and Toll-like receptor 4 (TLR4) can synergistically enhance the immune response. To investigate this synergy, we synthesized and evaluated a series of conjugated NOD2/TLR4 dual agonists comprising our in-house NOD2 agonist and two structurally distinct TLR4 agonists connected via flexible or rigid linkers. Our findings indicate that dual agonist activity toward both NOD2 and TLR4 is diminished upon conjugation. We also show that the linker chemistry significantly influences the kinetic solubility of these conjugates. Furthermore, the conjugates elicit distinct immunomodulatory effects in human primary peripheral blood mononuclear cells, characterized by a Th2-polarized cytokine response. These results provide insights into the structure-activity relationship of conjugated NOD2/TLR4 agonists and offer preliminary guidelines for tuning their solubility profiles.
新型免疫增强剂对于深化我们对免疫受体相互作用的理解以及应对传染病至关重要。先前的研究表明，含核苷酸结合寡聚化结构域蛋白2（NOD2）和Toll样受体4（TLR4）的共激活可协同增强免疫反应。为了研究这种协同作用，我们合成并评估了一系列共轭NOD2/TLR4双激动剂，这些双激动剂由我们内部的NOD2激动剂和通过柔性或刚性接头连接的两种结构不同的TLR4激动剂组成。我们的研究结果表明，共轭后对NOD2和TLR4的双激动剂活性均降低。我们还表明，接头化学性质显著影响这些共轭物的动力学溶解度。此外，这些共轭物在人原代外周血单核细胞中引发不同的免疫调节作用，其特征为Th2极化的细胞因子反应。这些结果为共轭NOD2/TLR4激动剂的构效关系提供了见解，并为调节其溶解度曲线提供了初步指导。
