Evaluation of In Vitro and In Vivo Anti-Tumor Effects and Pharmacokinetics of an Aniline–Pyrimidine Derivative B9

Non-small cell lung cancer (NSCLC) ranks among the malignancies with the highest global incidence and mortality rates. This study focused on a previous reported aniline–pyrimidine derivative, B9, which exhibited an IC50 of 21.43 μM against A549 cells. Herein, we further investigated its anti-NSCLC efficacy both in vitro and in vivo. The anti-proliferation assay proved B9 inhibits A549 cell proliferation in a concentration- and time-dependent manner, indicating B9 may have a good long-term effect in inhibiting the growth and division of cancer cells. The hemolysis assay revealed that B9 is non-hemolytic, with no observable hemolytic activity toward red blood cells, thereby confirming its preliminary biocompatibility. An in vivo efficacy study revealed that B9 effectively suppresses tumor growth within a low safe concentration of 2 μM in zebrafish models, confirming the anti-tumor efficacy of B9 was well preserved in animal models. Pharmacokinetic profiling showed B9 possesses unique metabolic profiles in rats and provides useful information of drug metabolism in vivo. Collectively, these findings establish B9 as a promising lead compound worthy of further development for NSCLC treatment.