Next, the cytotoxicity of the compound was evaluated in HeLa, H520 (lung cancer), and BEAS-2B (normal lung epithelial) cells using the MTT assay. As shown in Figure , the bismacrocycle 5 exhibited significant cytotoxicity across all cell lines at a minimal concentration of 3 μM after 24 hours of incubation. The half-maximal inhibitory concentration (IC50) values for HeLa, H520, and BEAS-2B cells were determined to be 3.700, 2.813, and 2.778 μM, respectively (Figure ). These relatively low IC50 values underscore the bismacrocycle's potent antitumor activity, which has not been previously reported in the literature. The bismacrocycle's potent cytotoxicity at low concentrations, with IC50 values notably lower than those of comparable nanohoops, suggests a superior therapeutic index and emphasizes its potential as an effective anticancer agent. This is particularly significant given the growing demand for drugs with high potency. Collectively, these findings lay a strong foundation for advancing the compound as a novel imaging agent and anticancer therapeutic, with promising applications in the imaging and treatment of various cancer types. Data are expressed as Mean ± SD, n = 3.