载阿霉素的槲皮素金属配合物靶向纳米粒的制备与表征

Preparation and characterization of doxorubicin-loaded quercetin metal complex targeting nanoparticles

摘要

目的: 制备载阿霉素的槲皮素金属配合物靶向递送系统,并考察其制剂学性质与抗肿瘤能力。方法: 以槲皮素为配体,通过调节锰铜摩尔比制备并筛选出最优双金属配合物载体,进一步调节阿霉素的投药量,筛选出最佳包封率和载药量的载药纳米粒,通过硫酸软骨素修饰制备靶向药物递送系统。对纳米粒进行颗粒粒径、Zeta电位、红外光谱、紫外光谱等理化性质表征;考察其体外释药行为、还原型谷胱甘肽(glutathione, GSH)分解能力、活性氧(reactive oxygen species,ROS)产生能力、血液相容性等;通过3T3和B16F10细胞系考察其安全性与抗肿瘤活性。结果: 本研究筛选出锰铜比为2∶1时合成的空白载体具有最佳抗肿瘤活性;当阿霉素投药量为0.05 mmol·L^－1时制备的纳米递送系统具有最佳载药量和包封率,靶向药物递送系统分别为(15.89±0.34)%和(90.21±0.45)%;最终经30%的硫酸软骨素修饰后制备靶向药物递送系统。各种表征表明本研究成功制备了双金属配合物靶向药物递送系统,粒径适宜且具有较高包封率和载药量。该递送系统在含GSH(10 mmol·L^－1)的pH 5.0条件下表现出较好的释药情况,具有良好的GSH分解能力、ROS产生能力、血液相容性、细胞相容性和肿瘤细胞毒性。结论: 本研究通过筛选金属比例、筛选投药量和修饰硫酸软骨素制备得到载阿霉素金属配合物的靶向药物递送系统,载药量高,具有良好的释药行为,这种使用槲皮素金属配合物与阿霉素的治疗策略通过化疗和化学动力学疗法协同抗肿瘤,可为协同促进抗肿瘤疗效提供新思路。

Abstract

Objective: To prepare a targeted delivery system of quercetin metal complex loaded with doxorubicin and to investigate its pharmaceutical properties and anti-tumor capability. Methods: With quercetin as the ligand, the optimal bimetallic complex carrier was prepared and screened by adjusting the molar ratios of manganese to copper, and the drug-loaded nanoparticles with the best encapsulation efficiency and drug loading were screened by further adjusting the dosage of doxorubicin. Finally, the targeted drug delivery system was prepared by chondroitin sulfate modification. The physicochemical properties of the nanoparticles such as particle size, Zeta potential, infrared spectrum and ultraviolet spectrum, were characterized. Then, the in vitro drug release behavior, glutathione (GSH) degradation capability, reactive oxygen species (ROS) production capability and blood compatibility were investigated. Finally, the safety and anti-tumor activity were investigated using 3T3 and B16F10 cell lines. Results: The blank carrier synthesized at the optimal manganese-copper ratio of 2∶1 exhibited the best anti-tumor activity. When the dosage of doxorubicin was 0.05 mmol·L^－1, the prepared nano-delivery system had the best drug loading and encapsulation efficiency, which were (15.89±0.34)% and (90.21±0.45)%, respectively. A targeted drug delivery system was then prepared after modification with 30% chondroitin sulfate. Various characterizations confirmed the successful preparation of the bimetallic complex targeted drug delivery system, with suitable particle size and high encapsulation efficiency and drug loading. The delivery system showed good drug release under pH 5.0 condition containing 10 mmol·L^－1 GSH, and exhibited favorable GSH degradation capability, ROS production capability, blood compatibility, cytocompatibility, and tumor cytotoxicity. Conclusion: In this study, a targeted drug delivery system loaded with doxorubicin metal complexes was successfully prepared by screening the metal ratio and doxorubicin dosage, followed by chondroitin sulfate modification. The system achieved high drug loading and favorable drug release behavior. This therapeutic strategy using quercetin metal complexes and doxorubicin exerts synergistic anti-tumor effects through combined chemotherapy and chemodynamic therapy, which may provide new ideas for enhancing synergistic anti-tumor efficacy.

关键词

槲皮素 / 阿霉素 / 金属配合物 / 硫酸软骨素 / 抗肿瘤

Key words

quercetin / doxorubicin / metal complex / chondroitin sulfate / anti-tumor

[1] YANG WL, SUN QH, ZHANG XD, et al. A novel doxorubicin/CTLA-4 blocker co-loaded drug delivery system improves efficacy and safety in antitumor therapy[J]. Cell Death Dis, 2024, 15(6): 386.
[2] 赵晨旭, 蔺雪峰, 韩轩茂. 小檗碱对阿霉素诱导心肌损伤的影响及保护效应[J]. 包头医学院学报, 2025, 41: 48-53, 89.
[3] DENG KL, YANG DX, ZHOU YP. Nanotechnology-based siRNA delivery systems to overcome tumor immune evasion in cancer immunotherapy[J]. Pharmaceutics, 2022, 14(7): 1344.
[4] LUAN F, HE XR, ZENG N. Tetrandrine: a review of its anticancer potentials, clinical settings, pharmacokinetics and drug delivery systems[J]. J Pharm Pharmacol, 2020, 72(11): 1491-1512.
[5] 王真玉, 龙巍然, 郑玉国, 等. 基于含氮羧酸配体/Co2+构筑的2D层状配合物的合成及结构表征[J]. 兴义民族师范学院学报, 2020(2): 120-124.
[6] SUN QQ, WANG Z, LIU B, et al. Recent advances on endogenous/exogenous stimuli-triggered nanoplatforms for enhanced chemodynamic therapy[J]. Coord Chem Rev, 2022, 451: 214267.
[7] CHEN G, YANG YY, XU Q, et al. Self-amplification of tumor oxidative stress with degradable metallic complexes for synergistic cascade tumor therapy[J]. Nano Lett, 2020, 20(11): 8141-8150.
[8] 王勤, 周刚, 申键. 槲皮素金属配合物合成及药理作用研究现状[J]. 北方药学, 2020, 17: 192-196.
[9] RONG Y, FAN ZX, YU ZJ, et al. An endoplasmic reticulum-targeting iridium(iii) complex induces immunogenic cell death in melanoma cells and enhances anti-PD-1 immunotherapy by remodeling tumor microenvironment[J]. Inorg Chem Front, 2023, 10(18): 5278-5291.
[10] ZHAO BL, BAN FZ, LI YH, et al. Exploiting mitochondrial dysfunction to overcome BRAF inhibitor resistance in advanced melanoma: the role of disulfiram as a copper ionophore[J]. Cell Death Dis, 2025, 16(1): 482.
[11] ZHANG Y, SHI JY, ZHU J, et al. Immunometabolic rewiring in macrophages for periodontitis treatment via nanoquercetin-mediated leverage of glycolysis and OXPHOS[J]. Acta Pharm Sin B, 2024, 14(11): 5026-5036.
[12] ROSTAMKHANI N, SALIMI M, ADIBIFAR A, et al. Enhanced anti-tumor and anti-metastatic activity of quercetin using pH-sensitive Alginate@ZIF-8 nanocomposites: in vitro and in vivo study[J]. Nanotechnology, 2024, 35(47): 1361-6528.
[13] LI L, ZHANG MX, LIU TT, et al. Quercetin-ferrum nanoparticles enhance photothermal therapy by modulating the tumor immunosuppressive microenvironment[J]. Acta Biomater, 2022, 154: 454-466.
[14] SAJID S, JAHAN N, HUMA Z, et al. Fabrication, characterization, and in vitro testing of quercetin-copper(II) complex[J]. Nano Biomed Eng, 2024, 16(3): 402-415.
[15] 齐帆, 赵晓平, 范小璇, 等. 中医药调控肿瘤微环境治疗脑胶质瘤的研究进展[J]. 中国实验方剂学杂志, 2024, 30: 303-314.
[16] WANG TT, FENG XC, LI LZ, et al. Effects of quercetin on tenderness, apoptotic and autophagy signalling in chickens during post-mortem ageing[J]. Food Chem, 2022, 383: 132409.
[17] 魏传宇, 于晓宇, 申倩, 等. 基于网络药理学消岩汤中夏枯草与白花蛇舌草治疗非小细胞肺癌作用机制的研究[J]. 中医肿瘤学杂志, 2021, 3: 31-41.
[18] AHN B, LEE SG, YOON HR, et al. Four-fold channel-nicked human ferritin nanocages for active drug loading and pH-responsive drug release[J]. Angew Chem Int Ed, 2018, 57(11): 2909-2913.
[19] ZENG XL, WANG YF, HAN JX, et al. Fighting against drug-resistant tumors using a dual-responsive Pt(IV)/Ru(II) bimetallic polymer[J]. Adv Mater, 2020, 32(43): e2004766.
[20] 魏素梅. 槲皮素衍生物的合成研究[D]. 济南: 山东师范大学, 2009.
[21] DE SOUZA RFV, DE GIOVANI WF. Antioxidant properties of complexes of flavonoids with metal ions[J]. Redox Rep, 2004, 9(2): 97-104.
[22] XU ZC, LUO QZ, HE YQ, et al. Endogenous nitric oxide releases in situ for RNS/ROS synergistic cancer therapy[J]. Adv Funct Materials, 2024, 34(17): 2314536.
[23] ZHANG L, SHEN H, LIU TT, et al. A pH/GSH dual-responsive triple synergistic bimetallic nanocatalyst for enhanced tumor chemodynamic therapy[J]. Small, 2025, 21(8): e2409836.
[24] LI AL, HUANG K, PAN WP, et al. Thiosemicarbazone mixed-valence Cu(I/II) complex against lung adenocarcinoma cells through multiple pathways involving cuproptosis[J]. J Med Chem, 2024, 67(11): 9091-9103.
[25] 雷西熙, 邓可斌, 姜锐. 槲皮素通过TLR4/IRAK4/NF-κB信号通路对过敏性鼻炎大鼠炎症反应的影响[J]. 中国药房, 2025, 36: 832-836.
[26] KONG HX, LI W, TIAN X, et al. NIR-activated FeCoOx nanozymes with tri-modal synergy for multi-target uproot drug-resistant bacterial keratitis[J]. Chem Eng J, 2025, 521: 166724.
[27] ZHOU H, BAO P, LIN YT, et al. Bimetallic nanoreactor mediates cascade amplification of oxidative stress for complementary chemodynamic-immunotherapy of tumor[J]. Biomaterials, 2025, 317: 123075.
[28] ZHENG NN, HU X, YAN L, et al. Bimetallic Cu@Ru core-shell structures with ligand effects for endo-exogenous stimulation-mediated dynamic oncotherapy[J]. Nano Lett, 2024, 24(20): 6165-6173.
[29] ZHU YY, HUANG CJ, ZHONG H, et al. A drug ligand based bimetallic coordination-driven self-assembly nanodrug for high-efficiency tumor chemodynamic therapy[J]. Mater Today Chem, 2024, 36: 101977.
[30] CHEN RF, GONG JL, YU ZY, et al. X-ray triggered bimetallic nanoassemblies as radiosensitizers and STING agonists for a CDT/radio-immunotherapy strategy[J]. Acta Biomater, 2025, 192: 366-376.
[31] ZHAO L, CHANG F, TONG Y, et al. A multifunctional bimetallic nanoplatform for synergic local hyperthermia and chemotherapy targeting HER2-positive breast cancer[J]. Adv Sci (Weinh), 2024, 11(16): e2308316.
[32] GONG F, CHEN MC, YANG NL, et al. Bimetallic oxide FeWOX nanosheets as multifunctional cascade bioreactors for tumor microenvironment-modulation and enhanced multimodal cancer therapy[J]. Adv Funct Materials, 2020, 30(49): 2002753.