Glycolytic metabolism enzymes have been implicated in the immunometabolism field through changes in metabolic status. PGK1 is a catalytic enzyme in the glycolytic pathway. Here, we set up a high-throughput screen platform to identify PGK1 inhibitors. DC-PGKI is an ATP-competitive inhibitor of PGK1 with an affinity of K_d = 99.08 nmol/L. DC-PGKI stabilizes PGK1 in vitro and in vivo, and suppresses both glycolytic activity and the kinase function of PGK1. In addition, DC-PGKI unveils that PGK1 regulates production of IL-1β and IL-6 in LPS-stimulated macrophages. Mechanistically, inhibition of PGK1 with DC-PGKI results in NRF2 (nuclear factor-erythroid factor 2-related factor 2, NFE2L2) accumulation, then NRF2 translocates to the nucleus and binds to the proximity region of Il-1β and Il-6 genes, and inhibits LPS-induced expression of these genes. DC-PGKI ameliorates colitis in the dextran sulfate sodium (DSS)-induced colitis mouse model. These data support PGK1 as a regulator of macrophages and suggest potential utility of PGK1 inhibitors in the treatment of inflammatory bowel disease.

通过代谢状态的变化，糖酵解代谢酶与免疫代谢领域有关。PGK1 是糖酵解途径中的催化酶。在这里，我们建立了一个高通量筛选平台来识别 PGK1 抑制剂。DC-PGKI 是一种 ATP 竞争性 PGK1 抑制剂，亲和力 Kd =  99.08 nmol/L _。DC-PGKI在体外和体内稳定 PGK1 ，并抑制 PGK1 的糖酵解活性和激酶功能。此外，DC-PGKI 揭示 PGK1 调节 IL-1 β的产生和 LPS 刺激的巨噬细胞中的 IL-6。从机制上讲，用 DC-PGKI 抑制 PGK1 会导致 NRF2（核因子-红细胞因子 2 相关因子 2，NFE2L2）积累，然后 NRF2 易位至细胞核并与Il-1β和Il-6基因的临近区域结合，并抑制 LPS 诱导的这些基因的表达。DC-PGKI 改善葡聚糖硫酸钠 (DSS) 诱导的结肠炎小鼠模型中的结肠炎。这些数据支持 PGK1 作为巨噬细胞的调节剂，并表明 PGK1 抑制剂在治疗炎症性肠病中的潜在用途。