| 陶世佳,赵乐梅,段威,等.二氧化硅负载酪氨酸酶纳米反应器的制备及其激活前药性能研究[J].中国海洋药物,2025,(6):-. |
| 二氧化硅负载酪氨酸酶纳米反应器的制备及其激活前药性能研究 |
| Preparation of Tyrosinase-Loaded Silica Nanoreactors and Evaluation of Their Ability for Activating Prodrugs |
| 投稿时间：2024-04-29 修订日期：2024-06-21 |
| DOI： |
| 中文关键词: 酶固定化 二氧化硅纳米反应器 催化动力学 催化纳米药物 前药催化 |
| English Keywords:Enzyme Immobilization Silica Nanoreactor Catalytic Kinetics Catalytic Nanomedicine Prodrug Catalysis |
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| 中文摘要: |
| 目的 天然酶易降解失活限制了其实际应用,二氧化硅纳米胶囊具有对酶的保护性和对反应底物的透过性,能够保证酶在应用中的稳定性和活性的发挥。目的 天然酶易降解失活限制了其实际应用，纳米载体将酶负载能够防止酶失活。制备负载酪氨酸酶的二氧化硅纳米反应器，并考察其激活前药的性能。方法 在不同pH条件下，通过检测催化反应的产物4-乙酰氨基-邻苯醌的吸光度值随时间的变化，筛选酶的最适pH，测定TYR的催化动力学参数Km和vmax。在最适pH下制备水相（含酶），通过反相（油包水）纳米乳液聚合技术合成载酪氨酸酶的二氧化硅纳米反应器。测定了纳米反应器的粒径、Zeta电位以及催化APAP的活性。结果 首先成功测定TYR对于APAP催化活性的最佳pH为7.4，测定了酪氨酸酶的催化动力学参数Km = 0.336 37 mol/L、vmax = 0.022 95 mmol/(L·min)。所制备的载酶二氧化硅纳米反应器粒径为 (115.2 ± 2.5) nm，Zeta电位为(?23.1 ± 2.1) mV，满足纳米药物的要求，具有良好的稳定性。负载酪氨酸酶的二氧化硅纳米反应器在体外表现出对APAP的优异催化活性。结论 基于二氧化硅纳米胶囊设计并制备负载酪氨酸酶的酶催化纳米反应器，具备良好的稳定性和生物活性，为实现前药激活疗法提供了新思路。 |
| English Summary: |
| Objective The susceptibility of natural enzymes to degradation and inactivation limits their practical application, and nanocarriers loaded with enzymes can prevent enzyme inactivation. Silica nanoreactors loaded with tyrosinase were prepared and examined for their performance in activating the precursor. Methods Under different pH conditions, the optimal pH of enzyme activity was screened by detecting the absorbance of the product of catalytic reaction 4-acetylamino-O-benzoquinone over time, and determining the values of catalytic kinetic parameters, Km and vmax. The aqueous phase (containing enzyme) was prepared at the optimal pH, and tyrosinase-loaded silica nanoreactors were synthesized by the inverse (water-in-oil) nanoemulsion technique. The particle size, Zeta potential, and activity of silica nanoreactors were determined. Results The optimal pH for the catalytic activity of TYR was firstly determined to be 7.4, and the catalytic kinetic parameters of tyrosinase, Km = 0.336 37 mol/L and vmax = 0.022 95 mmol/(L·min), were determined. The particle size of prepared enzyme-loaded silica nanoreactor was (115.2 ± 2.5) nm and the Zeta potential was (?23.1 ± 2.1) mV, which meet the requirement of nanomedicines with good stability. The tyrosinase-loaded silica nanoreactors successfully demonstrated high catalytic activity for APAP in vitro. Conclusion The design and preparation of enzyme-catalyzed nanoreactors loaded with tyrosinase based on silica nanocapsules with good stability and bioactivity provides a new idea to achieve prodrug activating therapy. |
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