Synthesis and Anticancer Evaluation of Amide‐Functionalized Calix[4]arene Derivatives: Induction of Apoptosis, Mitochondrial Dysfunction, and Inhibition of Cell Migration

ABSTRACT

Two amide‐functionalized calix[4]arene derivatives, namely 25,27‐bis{[(2‐((2‐aminoethyl)amino)ethyl)amino]carbonylmethoxy}‐26,28‐dihydroxycalix[4]arene ( 4a ) and 25,27‐bis[(2‐morpholinoethyl)aminocarbonylmethoxy]‐26,28‐dihydroxycalix[4]arene ( 4b ), were synthesized from 25,27‐diethoxycarbonylmethoxy‐26,28‐dihydroxycalix[4]arene (3) and structurally characterized by FT‐IR, ¹ H‐NMR, and elemental analysis. The anticancer activity of the compounds was evaluated in A549, Caco‐2, and LNCaP cancer cell lines using the MTT assay. Both derivatives exhibited dose‐dependent cytotoxicity. Compound 4a showed higher potency, with IC ₅₀ values of 15.12 ± 2.34 µM (A549), 17.31 ± 0.62 µM (Caco‐2), and 58.32 ± 1.17 µM (LNCaP), whereas compound 4b showed IC ₅₀ values of 61.51 ± 1.62 µM, 57.18 ± 3.72 µM, and 62.76 ± 2.95 µM, respectively. Further analyses revealed that the compounds significantly induced apoptosis, accompanied by mitochondrial membrane potential disruption and modulation of apoptosis‐related genes, including upregulation of BAX, CASP3, CASP8, CASP9, and P53, and downregulation of BCL2. In addition, the derivatives interfered with cell cycle progression, suppressed colony formation, and inhibited cell migration. The present study demonstrates that amide‐functionalized calix[4]arene derivatives exhibit multifaceted biological activity, supporting their further investigation as potential anticancer candidates.