Human papillomavirus‐16 E6‐positive cervical cancer attenuated by potent 2‐(4‐biphenylyl)‐N‐(1‐ethyl‐4‐piperidinyl) acetamide second‐generation analogs with improved binding affinity

Abstract Human papillomavirus (HPV) infection, particularly HPV16, is a major contributor to the development of cervical cancer. Given the urgent need for novel therapeutic strategies targeting HPV‐associated cancers, this study focuses on characterizing second‐generation analogs of a lead compound, as a potential inhibitor of HPV16‐E6. Protein–ligand docking, Gibbs binding free energy estimation, and molecular dynamics simulations were conducted. HPV16‐infected SiHa and CaSki cell lines were used. MTT (3‐(4,5‐dimethylthiazolyl‐2)‐2,5‐diphenyltetrazolium bromide) assay for proliferation and flow cytometry for target inhibition and apoptosis were employed. Computational and cell proliferation analyses revealed that modifications to E6‐855, particularly in the piperidinyl group, enhanced binding affinities against HPV16‐E6, with E6‐272 demonstrating superior binding properties. Molecular dynamics simulations confirmed the stable binding of E6‐272 to HPV16‐E6, supported by favorable binding energy estimates. E6‐272 inhibited the proliferation of SiHa and CaSki cells with GI 50 values of 32.56 and 62.09 nM, respectively. The compound reduced HPV16‐E6‐positive population, while inducing the early and late phase apoptosis in these cells. Structural alterations at the piperidinyl group of E6‐855 identified E6‐272 as a promising inhibitor of HPV16‐E6 with improved efficacy against HPV16‐E6. Further experimental validation of E6‐272 and its analogs warrant to advance its clinical utility in combating HPV‐associated cancers.