A Novel NQO1 Enzyme-Responsive Polyurethane Nanocarrier for Redox-Triggered Intracellular Drug Release.
A Novel NQO1 Enzyme-Responsive Polyurethane Nanocarrier for Redox-Triggered Intracellular Drug Release.
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DOI：
10.1021/acs.biomac.3c00134
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发表时间：
2023-04
影响因子：
6.2
通讯作者：
Jinhai Xie;Shuangyu Tian;Hanning Zhang;Congshu Feng;Yingchao Han;Honglian Dai;Lesan Yan
中科院分区：
文献类型：
--
作者：
Jinhai Xie;Shuangyu Tian;Hanning Zhang;Congshu Feng;Yingchao Han;Honglian Dai;Lesan Yan
近年来，设计响应肿瘤微环境刺激的纳米药物递送载体已成为发展癌症治疗的一个重要方面。其中，酶反应型纳米给药系统尤其有效，因为它利用肿瘤特异性和高表达的酶作为精确靶点，增加了药物在靶点的释放，减少了非特异性释放，提高了疗效，同时最大限度地减少了对正常组织的毒副作用。NAD(P)H：醌氧化还原酶1 （NQO1）是一种与癌症相关的重要还原酶，在一些癌细胞中过度表达，尤其是在肺癌和乳腺癌中。因此，设计对NQO1具有高选择性和高响应性的纳米载体对肿瘤的诊断和治疗具有重要意义。有报道称，在生理条件下，NQO1可以通过双电子还原特异性地还原三甲基锁定的苯醌结构，从而通过酶促反应快速内酯化。在此基础上，以二异氰酸酯、还原敏感单体TMBQ和聚乙二醇为共聚物，设计并合成了一种新型还原敏感聚氨酯嵌段共聚物（PEG-PTU-PEG）。通过核磁共振（1H NMR）和凝胶渗透色谱（GPC）验证了单体和聚合物的成功合成。然后，通过自组装成功制备了PEG-PTU-PEG胶束，并通过动态光散射（DLS）、1H NMR和GPC验证了其在Na2S2O4存在下的还原解离行为。然后，用微乳液法将模型药物多柔比星（DOX）包裹在该聚氨酯胶束的疏水核心中。结果表明，载药胶束还能实现氧化还原反应，快速释放被包被物质。体外细胞实验表明，peg - pta - peg胶束具有良好的生物相容性和较低的溶血率（<5%）。此外，在NQO1酶抑制剂（双oumarol）存在的情况下，通过荧光显微镜和流式细胞术检测，在A549和4T1细胞中观察到较低的药物从胶束释放，但在NIH-3T3对照细胞中没有。可以预见的是，在NQO1酶抑制剂存在的情况下，负载dox的胶束在4T1细胞中也表现出较低的细胞毒性。这些结果表明，在NQO1酶存在的还原环境下，载药聚氨酯胶束可以实现药物的特异性释放。因此，本研究为构建具有精准靶向和还原释放功能的聚氨酯纳米载体提供了新的选择，有利于肿瘤细胞内药物特异性释放和精准治疗。
The design of nano-drug delivery vehicles responsive to tumor microenvironment stimuli has become a crucial aspect in developing cancer therapy in recent years. Among them, the enzyme-responsive nano-drug delivery system is particularly effective, as it utilizes tumor-specific and highly expressed enzymes as precise targets, leading to increased drug release at the target sites, reduced nonspecific release, and improved efficacy while minimizing toxic side effects on normal tissues. NAD(P)H:quinone oxidoreductase 1 (NQO1) is an important reductase associated with cancer and is overexpressed in some cancer cells, particularly in lung and breast cancer. Thus, the design of nanocarriers with high selectivity and responsiveness to NQO1 is of great significance for tumor diagnosis and treatment. It has been reported that under physiological conditions, NQO1 can specifically reduce the trimethyl-locked benzoquinone structure through a two-electron reduction, resulting in rapid lactonization via an enzymatic reaction. Based on this, a novel reduction-sensitive polyurethane (PEG-PTU-PEG) block copolymer was designed and synthesized by copolymerizing diisocyanate, a reduction-sensitive monomer (TMBQ), and poly(ethylene glycol). The successful synthesis of monomers and polymers was verified by nuclear magnetic resonance (1H NMR) and gel permeation chromatography (GPC). Then, the PEG-PTU-PEG micelles were successfully prepared by self-assembly, and their reductive dissociation behavior in the presence of Na2S2O4 was verified by dynamic light scattering (DLS), 1H NMR, and GPC. Next, the model drug doxorubicin (DOX) was encapsulated into the hydrophobic core of this polyurethane micelles by microemulsion method. It was observed that the drug-loaded micelles could also achieve a redox response and rapidly release the encapsulated substances. In vitro cell experiments demonstrated that PEG-PTU-PEG micelles had good biocompatibility and a low hemolysis rate (<5%). Furthermore, in the presence of an NQO1 enzyme inhibitor (dicoumarol), lower drug release from micelles was observed in A549 and 4T1 cells by both fluorescence microscopy and flow cytometry assays, but not in NIH-3T3 control cells. Predictably, DOX-loaded micelles also showed lower cytotoxicity in 4T1 cells in the presence of NQO1 enzyme inhibitors. These results indicate that drug-loaded polyurethane micelles could accomplish specific drug release in the reducing environment in the presence of NQO1 enzymes. Therefore, this study provides a new option for the construction of polyurethane nanocarriers for precise targeting and reductive release, which could benefit the intracellular drug-specific release and precision therapy of tumors.