Development of Surface Chemical Strategies for Synthesizing Redox‐Responsive Diatomite Nanoparticles as a Green Platform for On‐Demand Intracellular Release of an Antisense Peptide Nucleic Acid Anticancer Agent

Abstract Redox‐responsive silica drug delivery systems are synthesized by aeco‐friendly diatomite source to achieve on‐demand release of peptide nucleic acid (PNA) in tumor reducing microenvironment, aiming to inhibit the immune checkpoint programmed cell death 1 receptor/programmed cell death receptor ligand 1 (PD‐1/PD‐L1) in cancer cells. The nanoparticles (NPs) are coated with polyethylene glycol chains as gatekeepers to improve their physicochemical properties and control drug release through the cleavable disulfide bonds (S–S) in a reductive environment. This study describes different chemical conditions to achieve the highest NPs’ surface functionalization yield, exploring both multistep and one‐pot chemical functionalization strategies. The best formulation is used for covalent PNA conjugation via the S–S bond reaching a loading degree of 306 ± 25 µg PNA mg −1 DNPs . These systems are used for in vitro studies to evaluate the kinetic release, biocompatibility, cellular uptake, and activity on different cancer cells expressing high levels of PD‐L1. The obtained results prove the safety of the NPs up to 200 µg mL −1 and their advantage for controlling and enhancing the PNA intracellular release as well as antitumor activity. Moreover, the downregulation of PD‐L1 observed only with MDA‐MB‐231 cancer cells paves the way for targeted immunotherapy.