Role of acetate/benzoate substituents on the anticancer activities, in vitro and in silico DNA/BSA interactions of new 1,4-disubstituted 1,2,3-triazole derivatives

• Novel 1,4-disubstituted 1,2,3-triazoles synthesized via CuAAC method. • X-ray crystallography confirmed triclinic P −1 space group for compound 3. • Compound 4 outperformed cisplatin against Caco-2 cancer cells. • Benzoate group enhances cytotoxicity and selectivity in cancer cells. • In-vitro and in-silico studies reveal binding mechanism to CT-DNA and BSA . We report herein two new 1,4-disubstituted 1,2,3-triazole derivatives which were synthesized by copper(I)-catalyzed azide alkyne cycloaddition (CuAAC). Their structures were elucidated by FTIR, 1 H NMR, 13 C APT NMR, MALDI-TOF-MS and elemental analysis. Moreover, X-ray crystallography studies of compound 3 demonstrated that the compound adapted triclinic system P -1 space group. Its asymmetric unit contains two crystallographically independent molecules. Hirshfeld surface analysis indicated that hydrogen bonding and van der Waals interactions dominate the crystal packing. The compounds 3 and 4 were investigated for their DNA/BSA interactions using in vitro and in silico techniques, showing that binding to the major groove of calf thymus DNA (CT-DNA) and bovine serum albumin (BSA) was mediated by polar and hydrophobic interactions. The anticancer activities of the compounds were tested by MTT assay on human cell lines, breast cancer (MDA-MB-231), prostate cancer (LNCaP), colorectal adenocarcinoma (Caco-2), and embriyonic kidney cell line (HEK-293) as a healty cell line. The compound 4 exhibited more cytotoxic activity than cisplatin on Caco-2 cancer cell line and demonstrated selectivity against MDA-MB-231 cells with an SI>2 value. Annexin V staining indicated apoptosis induction via loss of mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) generation in MDA-MB-231 cells. These findings suggest that the benzoate group positively affects the biological activities of these compounds.