Department of Biomedical Engineering, University of Minnesota, Minneapolis, Minnesota 55455, United States.
Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota 55455, United States.
Mol Pharm. 2023 Apr 3;20(4):1884-1897. doi: 10.1021/acs.molpharmaceut.2c00385. Epub 2023 Mar 10.
Tumor necrosis factor (TNF) is a key regulator of immune responses and plays a significant role in the initiation and maintenance of inflammation. Upregulation of TNF expression leads to several inflammatory diseases, such as Crohn's, ulcerative colitis, and rheumatoid arthritis. Despite the clinical success of anti-TNF treatments, the use of these therapies is limited because they can induce adverse side effects through inhibition of TNF biological activity, including blockade of TNF-induced immunosuppressive function of TNFR2. Using yeast display, we identified a synthetic affibody ligand (ABY) with high binding affinity and specificity for TNFR1. Functional assays showed that the lead affibody potently inhibits TNF-induced NF-κB activation (IC of 0.23 nM) and, crucially, does not block the TNFR2 function. Additionally, ABY acts non-competitively─it does not block TNF binding or inhibit receptor-receptor interactions in pre-ligand-assembled dimers─thereby enhancing inhibitory robustness. The mechanism, monovalent potency, and affibody scaffold give this lead molecule uniquely strong potential as a therapeutic candidate for inflammatory diseases.
肿瘤坏死因子（TNF）是免疫反应的关键调节剂，在炎症的启动和维持中起着重要作用。TNF 表达的上调导致了几种炎症性疾病，如克罗恩病、溃疡性结肠炎和类风湿性关节炎。尽管抗 TNF 治疗在临床上取得了成功，但这些疗法的使用受到限制，因为它们通过抑制 TNF 的生物活性，包括阻断 TNF 诱导的 TNFR2 的免疫抑制功能，可能会引起不良反应。我们使用酵母展示技术，鉴定了一种对 TNFR1 具有高结合亲和力和特异性的合成亲和体配体（ABY）。功能分析表明，该先导亲和体能够强烈抑制 TNF 诱导的 NF-κB 激活（IC 为 0.23 nM），并且关键是不阻断 TNFR2 的功能。此外，ABY 非竞争性地发挥作用——它不阻断 TNF 的结合或抑制预配体组装的二聚体中的受体-受体相互作用——从而增强了抑制的稳健性。该先导分子的作用机制、单价效力和亲和体支架使其具有作为炎症性疾病治疗候选药物的独特强大潜力。
