Glycosylated Shikonin derivative Z-1-2 inhibits triple-negative breast Cancer growth and metastasis through glycerophospholipid metabolism modulation

This study developed a novel glycosylated shikonin derivative ((2 S ,3 R ,4 S ,5 R ,6 R )-2-(Acetamido)-6-(3-(1,4-dihydroxy-2-(1-hydroxy-4-methylpent-3-en-1-yl)naphthalen-5-yl)oxy-3-oxopropyl)oxy-4,5-dihydroxy-tetrahydro-2H-pyran-3-yl acetate, Z-1-2) by introducing a 2-deoxy-2-aminoglycoside moiety via succinic anhydride linkage to enhance its anti-tumor activity against triple-negative breast cancer. Z-1-2 exhibited superior cytotoxicity to parent shikonin in MDA-MB-231 cells (IC 50 of 4.05 ± 0.11 μM vs. 5.06 μM), with minimal toxicity to normal endothelial cells (HUVEC IC 50 = 2.93 μM). Mechanistic investigations revealed that Z-1-2 suppressed cell migration (wound healing inhibition at 8 μM, p < 0.001) and induced apoptosis (annexin V+ cells dose-dependently increased by 28–53%) through the modulation of glycerophospholipid metabolism, alanine-glutamate homeostasis, and riboflavin cycling pathways. In a model of triple-negative breast cancer using xenografts, Z-1-2 demonstrated dose-dependent tumor inhibition (58.7% at 5 mg/kg vs. control, p < 0.05), with improved safety profiles compared with those of shikonin. Untargeted metabolomics identified lysoPC 18:0 and phosphatidylcholine (P-16:0/0:0) as key biomarkers linked to membrane fluidity disruption and Src kinase inhibition. This work highlights Z-1-2 as a promising candidate for triple-negative breast cancer therapy, combining improved pharmacokinetics with multi-target metabolic modulation. • A novel glycosylated shikonin derivative Z-1-2 was synthesized by introducing 2-deoxy-2-aminoglycoside via succinic anhydride linkage. • Z-1-2 exhibited enhanced cytotoxicity against TNBC MDA-MB-231 cells (IC₅₀ = 4.05 ± 0.11 μM) and reduced toxicity vs. parent shikonin, with dose-dependent anti-migration and pro-apoptosis effects in vitro. • In TNBC xenograft models, Z-1-2 achieved 58.7% tumor inhibition at 5 mg/kg with favorable in vivo safety and no obvious body weight loss in mice. • Untargeted metabolomics identified glycerophospholipid metabolism as the core regulatory pathway, with lysoPC 18:0 and PC (P-16:0/0:0) as key biomarkers for Z-1-2's anti-TNBC activity. • Z-1-2 modulates multiple metabolic pathways (glycerophospholipid, alanine-glutamate, riboflavin cycling) and disrupts membrane fluidity/Src kinase activity, serving as a promising multi-target TNBC therapeutic candidate.