将蛋白水解酶固定在纳米载体上可通过降解致密的细胞外基质（ECM）来有效改善肿瘤中的药物扩散。在此，探索了透明质酸酶、菠萝蛋白酶和胶原酶 (Coll) 在介孔二氧化硅纳米粒子 (MSN) 上的固定和释放行为。合成了一系列具有大且可调孔径的阳离子MSN（CMSN），并与两种不同孔径的阴离子MSN一起进行了研究。 CMSNs4.0对透明质酸酶和菠萝蛋白酶的酶负载能力最高，CMSNs4.5对Coll的酶负载能力最好。高静电相互作用、匹配的孔径以及大的孔体积和表面积有利于固定化。酶构象和表面电荷随 pH 值的变化、固定化酶周围空间的存在以及孔结构的深度都会影响释放率和可调性。最佳的 CMSNs-酶复合物表现出对胰腺肿瘤的深度和均匀渗透，这是一种具有最致密 ECM 的肿瘤模型，其中 CMSNs4.5-Coll 是最好的。负载阿霉素 (DOX) 后，CMSNs-酶复合物诱导高抗肿瘤效率。可以想象，DOX/CMSNs4.5-NH2-Coll纳米药物表现出最有效的肿瘤治疗效果，肿瘤生长抑制率为86.1%。该研究提供了优异的纳米载体-酶复合物，并为增强肿瘤渗透和治疗提供了指导性理论。版权所有 © 2024。由 Elsevier B.V. 出版。

Immobilization of proteolytic enzymes onto nanocarriers is effective to improve drug diffusion in tumors through degrading the dense extracellular matrix (ECM). Herein, immobilization and release behaviors of hyaluronidase, bromelain, and collagenase (Coll) on mesoporous silica nanoparticles (MSNs) were explored. A series of cationic MSNs (CMSNs) with large and adjustable pore sizes were synthesized, and investigated together with two anionic MSNs of different pore sizes. CMSNs4.0 exhibited the highest enzyme loading capacity for hyaluronidase and bromelain, and CMSNs4.5 was the best for Coll. High electrostatic interaction, matched pore size, and large pore volume and surface area favor the immobilization. Changes of the enzyme conformations and surface charges with pH, existence of a space around the immobilized enzymes, and the depth of the pore structures, affect the release ratio and tunability. The optimal CMSNs-enzyme complexes exhibited deep and homogeneous penetration into pancreatic tumors, a tumor model with the densest ECM, with CMSNs4.5-Coll as the best. Upon loading with doxorubicin (DOX), the CMSNs-enzyme complexes induced high anti-tumor efficiencies. Conceivably, the DOX/CMSNs4.5-NH2-Coll nanodrug exhibited the most effective tumor therapy, with a tumor growth inhibition ratio of 86.1 %. The study provides excellent nanocarrier-enzyme complexes, and offers instructive theories for enhanced tumor penetration and therapy.Copyright © 2024. Published by Elsevier B.V.