Enzyme-Activated Cascade Biocatalysis of a Hyaluronic Acid-Engineered Biomineralization Nanozyme for Amplified Multimodal Cancer Treatment

Abstract The integration of nanozyme-based catalytic activity interventions within the tumor microenvironment (TME) with phototherapies represents a promising approach for triggering multimodal synergistic cancer therapy. Herein, a protein-mineralized nanozyme was fabricated via in situ biomineralization, utilizing iron ions coordinated with glucose oxidase (GOx-Fe). Unlike traditional nanozyme systems that rely on passive targeting or a single catalytic function, the design uniquely integrates a cascade therapeutic system within a single biocompatible platform. This nanozyme possesses dual catalytic properties to facilitate glucose oxidation and the conversion of hydrogen peroxide (H2O2) into hydroxyl radicals (•OH) and oxygen (O2). This dual-catalytic mechanism establishes a self-augmenting cascade that concurrently activates starvation therapy (ST) and chemodynamic therapy (CDT) within a signal nanosystem. To achieve tumor-specific delivery, the GOx-Fe nanozyme was modified with a polymer consisting of Chlorin e6-functionalized PEGylated hyaluronic acid (mHDC). The resulting GOx-Fe@mHDC nanozyme exhibits a uniform spherical morphology with an average hydrodynamic diameter of approximately 50 nm and a negative surface charge of about −19 mV. It maintains stability across diverse physiological environments but undergoes activation in the enzyme-rich TME, ensuring targeted and selective therapeutic effects. The nanozyme demonstrates efficient glucose consumption (approaching the performance of free GOx) and significant intracellular H2O2 generation (290% higher than free GOx), leading to enhanced •OH production and O2 self-supply. In A549 tumor-bearing mice, intravenous administration of GOx-Fe@mHDC followed by 660 nm laser irradiation resulted in marked tumor growth inhibition (tumor weight suppression rate of 86%) with negligible systemic toxicity. Importantly, the developed GOx-Fe@mHDC system exhibited strong catalytic stability, targeted tumor accumulation, effective endolysosomal escape, selective cytotoxicity, and substantial in vivo tumor suppression with excellent biosafety. This research introduces a versatile biomineralization technique for constructing Fe-based nanozymes with dual catalytic and tumor-specific capabilities, offering a scalable approach for advancing cancer therapeutics.