Self-preparation system using glucose oxidase-inspired nitroreductase amplification for cascade-responsive drug release and multidrug resistance reversion

Abstract

Early antitumor therapy is an important determinant of survival in patients with cancer. Utilization of specific pathological states, such as hypoxia, greatly promotes the development of intelligent drug delivery systems (DDSs) for targeted antitumor therapy. However, a slight decrease in oxygen levels in early-stage tumors is not sufficient to trigger hypoxia-responsive drug release. Nitroreductase (NTR) is overexpressed in bioreductive hypoxic cancers, and its expression level has been verified to be directly related to hypoxic status. Herein, using glucose oxidase (GOx) as an O₂-consuming agent to exacerbate hypoxia, a cascade strategy of GOx-induced overexpression of NTR and amplified NTR-catalyzed release was proposed for early antitumor therapy. Briefly, NTR-sensitive p-nitrobenzyl chloroformate (PNZ-Cl) was adopted to conjugate with the polysaccharide chitosan (CS) and self-assemble into CS-PNZ-Cl micelles. These polymer micelles possess the dual abilities to specifically immobilize GOx and load mitoxantrone (MIT) to form the NTR-responsive nanocascade reactor GOx/MIT@CS-PNZ-Cl. First, as a "key", tumor hypoxia triggers the initial release of GOx, which serves as the O₂-consuming agent when catalyzing its reaction with glucose, which is accompanied by H₂O₂ production. Depleted oxygen levels facilitate the expression of NTR, which in turn amplifies the capacity of the nanocascade reactor to decompose into secondary micelles for enhanced intratumoral permeation. GOx-inspired NTR amplification further elicits MIT release, realizing a synergistic "domino effect" cascade. In addition, upregulated H₂O₂ has been shown to effectively reverse GSH-mediated MIT resistance, reaching the superior tumor inhibition rate of 93.08%. This GOx-based NTR-responsive nanocascade reactor provides amplification of the bioreductive hypoxic tumor microenvironment for early antitumor therapy.

Keywords: Bioreductive hypoxic tumor microenvironment; Cascade nitroreductase-responsive release; Glucose oxidase; Mitoxantrone.