GSH/ROS Dual-Responsive Supramolecular Nanoparticles Based on Pillar[6]arene and Betulinic Acid Prodrug for Chemo–Chemodynamic Combination Therapy
Abstract
:1. Introduction
2. Results and Discussion
2.1. Synthesis of the Host WP6 and Guest BA-G
2.2. Fabrication of Supramolecular Nanoparticles: BNPs and GOx@BNPs
2.3. Drug Release Behavior
2.4. GOx@BNPs Induced Cascade Reactions
2.5. Cytotoxicity of GOx@BNPs
2.6. Cellular Internalization and Subcellullar Distribution
2.7. Mechanism of Cancer Inhibition
2.7.1. Elevation of ROS
2.7.2. Decrease in ATP
2.7.3. Evaluation of Mitochondrial Transmembrane Potential (MTP)
2.7.4. Induction of Apoptosis
3. Materials and Methods
3.1. General Information
3.2. Preparation of BA-G
3.2.1. Synthesis of Compound 3
3.2.2. Synthesis of Compound 5
3.2.3. Synthesis of Compound 8
3.2.4. Synthesis of Compound 9
3.2.5. Synthesis of Copound BA-G
3.3. Preparation of BNPs and GOx@BNPs
where mGOx-loaded, mGOx, mWP6, and mBA-G are the mass of GOx encapsulated into the vesicles and the mass of GOx, WP6, and BA-G added, respectively. The mass of GOx was measured via fluorescence spectroscopy at 520 nm and calculated according to a standard calibration curve of GOx in deionized water, with concentrations from 10 to 100 μg/mL.Loading content (wt %) = (mGOx-loaded/mGOx-loaded + mWP6 + mBA-G) × 100Encapsulation efficiency (%) = (mGOx-loaded/mGOx) × 100
3.4. Synthesis of FITC-GOx
3.5. In Vitro Release of BA
3.6. pH Variation under Different Concentrations of Glucose
3.7. Measurement of ·OH Generation
3.8. In Vitro Cytotoxicity by Intracellular Drug Delivery
3.9. Live/Dead Cell Staining
3.10. Cellular Uptake and Subcellular Distribution
3.11. Intracellular Reactive Oxygen Species (ROS) Detection
3.12. Intracellular ATP Level Measurements
3.13. Evaluation of Mitochondrial Transmembrane Potential (MTP)
3.14. Flow Cytometric Analysis of Apoptosis
3.15. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Zhu, P.; Luo, W.; Qian, J.; Meng, C.; Shan, W.; Xu, Z.; Zhang, W.; Liu, X.; Ling, Y. GSH/ROS Dual-Responsive Supramolecular Nanoparticles Based on Pillar[6]arene and Betulinic Acid Prodrug for Chemo–Chemodynamic Combination Therapy. Molecules 2021, 26, 5900. https://doi.org/10.3390/molecules26195900
Zhu P, Luo W, Qian J, Meng C, Shan W, Xu Z, Zhang W, Liu X, Ling Y. GSH/ROS Dual-Responsive Supramolecular Nanoparticles Based on Pillar[6]arene and Betulinic Acid Prodrug for Chemo–Chemodynamic Combination Therapy. Molecules. 2021; 26(19):5900. https://doi.org/10.3390/molecules26195900
Chicago/Turabian StyleZhu, Peng, Weidan Luo, Jianqiang Qian, Chi Meng, Wenpei Shan, Zhongyuan Xu, Wei Zhang, Xin Liu, and Yong Ling. 2021. "GSH/ROS Dual-Responsive Supramolecular Nanoparticles Based on Pillar[6]arene and Betulinic Acid Prodrug for Chemo–Chemodynamic Combination Therapy" Molecules 26, no. 19: 5900. https://doi.org/10.3390/molecules26195900