Drug Screening for Glycolysis Pathway in Living Cancer Cells Using 19F NMR

Glycolysis is a fundamental process for the generation of cellular energy, and its dysregulation has been linked to a number of diseases, including obesity, neurological disorders, and cancer. Targeting glycolysis is therefore a promising therapeutic strategy. However, effective drugs that specifically target glycolysis are still lacking. In this study, we introduce a novel approach utilizing ¹⁹F NMR to monitor early glycolysis in living MCF-7 cells. By tracking metabolites downstream of the glucose analogue 2-fluoro-2-deoxyglucose (2-FDG), we successfully observed the activity of key glycolytic components, such as glucose transporters (GLUTs) and hexokinase (HKs). Our results reveal distinct metabolic profiles upon inhibition of these targets, advancing the understanding of glycolytic regulation. In addition, we applied this approach to screen traditional Chinese medicines for their effects on glycolysis and identified Salvia miltiorrhiza and Fructus evodiae as modulators with contrasting effects on glycolytic metabolism. This dual modulation highlights their potential as valuable tools for therapeutic intervention. Our study provides an innovative methodology for both the exploration of glycolytic pathways and the discovery of novel therapeutics, offering new perspectives for drug development targeting metabolic diseases.

糖酵解（Glycolysis）是细胞能量生成的核心途径，其失调与肥胖、神经系统疾病及癌症等多种疾病密切相关。因此，靶向糖酵解成为极具潜力的治疗策略。然而，目前仍缺乏能够特异性靶向糖酵解的有效药物。本研究提出一种全新方法，利用¹⁹F核磁共振（¹⁹F NMR）监测活MCF-7细胞内的早期糖酵解过程。通过追踪葡萄糖类似物2-氟-2-脱氧葡萄糖（2-fluoro-2-deoxyglucose，2-FDG）下游的代谢物，我们成功观测到葡萄糖转运蛋白（glucose transporters, GLUTs）与己糖激酶（hexokinase, HKs）等关键糖酵解组分的活性。研究结果显示，抑制上述靶点后会呈现出独特的代谢谱特征，加深了我们对糖酵解调控机制的理解。此外，我们将该方法应用于中药的糖酵解调控效应筛选，最终鉴定出丹参（Salvia miltiorrhiza）和吴茱萸（Fructus evodiae）这两种对糖酵解代谢具有相反调控作用的调节剂。这种双向调控特性凸显了它们作为治疗干预潜在工具的应用价值。本研究为糖酵解通路的探索及新型治疗药物的发现提供了创新性方法，也为靶向代谢性疾病的药物研发开辟了新视角。