The xenograft model mice (n = 6 per group) were administered DDO-5936 intravenously at 100 mg/kg. Blood samples with a volume of 150 μl were collected into heparinized Eppendorf tubes at indicated time points (0, 0.25, 0.5, 1, 2, 8, 12, and 24 hours) and centrifuged at 3000 rpm for 10 min to obtain plasma samples (the samples were stored at −80°C until liquid chromatography–tandem mass spectrometry analysis). Tumor samples were collected at the same time points before homogenized. The concentration of the protein lysates of tumor tissue was determined by a bicinchoninic acid kit. Then, the protein lysates were analyzed by Western blots.

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