6H2L, a novel derivative of bifendate, dually suppresses glycolysis and glutaminolysis in hepatocellular carcinoma via inhibiting YAP stability.

Hepatocellular carcinoma (HCC) has poor clinical outcomes due to limited chemotherapy response and low targeted therapy efficacy. Metabolic reprogramming, a key hallmark of HCC tumorigenesis, represents a promising therapeutic target. This study elucidated the regulatory effects and underlying mechanism of 6H2L, a novel bifendate derivative, on metabolic reprogramming in HCC. CCK-8, EdU, and colony formation assays were employed to evaluate cell proliferation. The expression of key enzymes involved in glycolysis and glutaminolysis was analyzed by western blot and qPCR. Cycloheximide/MG-132 treatment, co-immunoprecipitation, and luciferase reporter assay were performed to investigate the effects of 6H2L on Yes associated protein (YAP) stability, localization, and transcriptional activity. The anti-tumor activity of 6H2L were verified by a subcutaneous xenograft model. 6H2L inhibited HCC cell proliferation by dually suppressing glycolysis and glutaminolysis. YAP was found to promote HCC growth in vivo by facilitating both glycolysis and glutaminolysis. Mechanistically, 6H2L directly bond to nuclear Dbf2-related kinase 1 (NDR1) and activated NDR signaling, thereby increasing phosphorylation of YAP at Ser127, which enhanced the binding of YAP to 14-3-3 protein and facilitated subsequent ubiquitin-mediated YAP degradation. This process inhibited YAP activity and reduced glycolysis and glutaminolysis. NDR1 knockdown attenuated the inhibitory effects of 6H2L on YAP stability, metabolic reprogramming, and cell proliferation. In vivo studies confirmed that 6H2L inhibited HCC growth via NDR/YAP axis with a favorable safety profile. Collectively, these findings identify the NDR/YAP axis as a novel target for disrupting glycolysis and glutaminolysis in HCC, and further highlight 6H2L as a promising therapeutic candidate.