Fig. 7.

Metabolomic profiling of compound 4j in human cells. a SW620 cells were treated with 4j (2.5 µM, 4 h) or DMSO and collected for metabolomic analysis. Untargeted metabolomics showed a number of significant metabolite changes (p < 0.01) measured in positive ionization mode. Bubble sizes represent fold change, while increasing statistical significance is represented by darker shadings. The two bubbles at RT ≈26 min correspond to hydrogen or sodium adduct of a cellular metabolite of 4j (M = 4j – C₅H₆BNO₃). Several species in the expanded inset were deduced to be potential phosphatidylcholines (PCs) by mass-to-charge ratio and retention time. b Multiple reaction monitoring of PC-related transitions (M+H→ phosphocholine) confirmed elevations in 26:0 PC in 4j-treated cells. c Negative mode fragmentation (M+HCO₂ ⁻→ RCOO⁻) revealed PC(26:0) to be a mixture of PC(10:0–16:0) and PC(12:0–14:0) species. For b and c, the data represent average values ± S.E.M. values from five independent experiments per group. Statistical significance was calculated with unpaired students t-tests comparing 4j-treated to DMSO-treated groups; ***p < 0.001; ****p < 0.0001