混合卤代亚酞菁的设计合成及其溶酶体靶向光动力治疗研究

doi:10.6023/A26040099

摘要/Abstract

光动力疗法(PDT)是前景广阔的肿瘤靶向治疗技术, 开发高性能光敏染料是其发展的核心关键. 亚酞菁(SubPc)作为性能优异的大环光敏染料, 在肿瘤PDT治疗中极具应用价值. 本研究通过一锅法制备四种氟氯梯度取代低对称亚酞菁(SubPc 4~7), 结合理论计算系统探究卤素取代对其光物理及抗肿瘤性能的调控规律. 结果表明, 随氯取代比例升高, 分子吸收与荧光波长持续红移; 荧光量子产率与单线态氧产率呈负相关, 多氯累加重原子效应可有效促进系间窜越, 提升光敏能力. 该系列分子具有良好水相光学稳定性, 可靶向肿瘤细胞溶酶体并产生活性氧. 体外实验证实SubPc 4肿瘤细胞杀伤率达82%, 小鼠模型及组织切片实验进一步验证其优异的体内PDT抑瘤效果. 本研究为亚酞菁靶向光动力抗肿瘤应用提供了重要实验依据.

关键词: 光动力疗法, 亚酞菁, 卤素取代, 单线态氧, 溶酶体靶向

Photodynamic therapy (PDT) is a promising targeted tumor treatment strategy, and developing high-performance photosensitizers is crucial for its advancement. As excellent macrocyclic photosensitive dyes, subphthalocyanines (SubPcs) show great application potential in tumor PDT. In this study, four low-symmetry SubPc derivatives with gradient fluoro-chloro substitution (SubPc 4~7) were prepared via a one-pot method. Combined with theoretical calculations, the regulatory effects of halogen substitution on their photophysical and antitumor properties were systematically investigated. The results showed that increasing chlorine substitution ratio led to progressive redshifts in both absorption and fluorescence emission wavelengths. Fluorescence quantum yield was negatively correlated with singlet oxygen quantum yield, and the cumulative heavy-atom effect of multiple chlorine atoms effectively promoted intersystem crossing, thereby enhancing photosensitization capability. These derivatives exhibited good optical stability in aqueous environments and could target lysosomes of tumor cells to generate reactive oxygen species. In vitro experiments confirmed that SubPc 4 achieved a tumor cell killing rate of 82%. In vivo mouse models and tissue section analysis further verified its potent antitumor efficacy in PDT. This study provides an important experimental basis for the application of SubPc derivatives in targeted photodynamic antitumor therapy.

Key words: photodynamic therapy, subphthalocyanine, halogen substitution, singlet oxygen, lysosome targeting

引用此文

张恩泽, 田济华, 沈一诺, 曾绍红, 王叶梅, 郑芬芬. 混合卤代亚酞菁的设计合成及其溶酶体靶向光动力治疗研究[J]. 化学学报, 2026, 84(9): 1442-1451.

Enze Zhang, Jihua Tian, Yinuo Shen, Shaohong Zeng, Yemei Wang, Fenfen Zheng. Design and Synthesis of Mixed Halogenated Subphthalocyanines for Lysosome-Targeted Photodynamic Therapy[J]. Acta Chimica Sinica, 2026, 84(9): 1442-1451.

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图/表 11

图式1. 化合物SubPcs 4~7的合成路线

Scheme 1. The synthesis route for compounds SubPcs 4~7

图1. (a)化合物SubPcs 4~7的紫外吸收光谱图; (b)化合物SubPcs 4~7的荧光发射光谱图

Figure 1. (a) The UV absorption spectra of compounds SubPcs 4~7; (b) The fluorescence emission spectra of compounds SubPcs 4~7

图2. 化合物SubPc 4 (a)、SubPc 5 (b)、SubPc 6 (c)和SubPc 7 (d)在二氯甲烷中的循环伏安图

Figure 2. Cyclic voltammograms of compounds SubPc 4 (a), SubPc 5 (b), SubPc 6 (c), and SubPc 7 (d) in dichloromethane

	E_ox/V	E_red/V			E_R^a	H-L (V)^b
	E_ox/V	1st	2nd	3rd	E_R^a	H-L (V)^b
4	1.77	－1.43	－1.60	－2.23	0.17	3.20
5	1.70	－1.40	－1.59	－2.20	0.19	3.10
6	1.70	－1.40	－1.69	－2.20	0.29	3.10
7	1.67	－1.40	－1.56	－2.18	0.16	3.07

表1. 化合物SubPcs 4~7在含有0.1 mol/L四丁基高氯酸铵(TBAP)的CH2Cl2中的半波电位(V vs. SCE)

Table 1. Half-wave potential (V vs. SCE) of SubPcs 4~7 in CH2Cl2 containing 0.1 mol/L TBAP

图3. F12-SubPc, Cl12-SubPc和SubPcs 4~7的MO能级图

Figure 3. MO energy level diagrams of F12-SubPc, Cl12-SubPc and SubPcs 4~7

图4. 4T1细胞摄取SubPcs 4~7 4 h的荧光图片, 比例尺为10 μm

Figure 4. Fluorescence images of SubPcs 4~7 uptake by 4T1 cells for 4 h, the scale bar is 10 μm

图5. (a) 以SubPc 4为光敏剂, DPBF在412 nm处的紫外吸收图; (b) SubPc 4单线态氧生成能力测试

Figure 5. (a) UV absorption spectrum of DPBF at 412 nm using SubPc 4 as photosensitizer; (b) Testing the singlet oxygen generation capability of SubPc 4

		氟原子数	氯原子数	Φ_△/%	Φ_F/%
4	Cl₁₀SubPc	—	10	59.29	17.25
5	Cl₈SubPc	—	8	41.56	19.99
6	F₄Cl₄SubPc	4	4	35.81	28.45
7	F₈Cl₂SubPc	8	2	19.73	32.59

表2. SubPcs 4~7中F、Cl原子数及对应的单线态氧产率(Φ△)和荧光量子产率(ΦF)

Table 2. F/Cl atom counts and the corresponding singlet-oxygen yield (Φ△) and fluorescence yield (ΦF) of SubPcs 4~7

图6. SubPcs 4~7在4T1细胞中580~590 nm激光照射后的DCFH染色的CLSM图像, 比例尺为50 µm

Figure 6. CLSM images of DCFH staining after 580~590 nm laser irradiation of SubPcs 4~7 in 4T1 cells, the scale bar is 50 μm

图7. (a) SubPc 4对4T1细胞的体外光毒性和暗毒性评价(MTT法). 细胞以不同浓度SubPc 4 (2.5、5.0、10.0、20.0、50.0 μg/mL)孵育24 h后, 光毒性组采用560~580 nm激光(0.5 W/cm2)照射10 min, 暗毒性组避光处理. 数据以均值±标准差表示(n=3). (b)根据图(a)中光毒性组剂量-效应数据拟合的半数抑制浓度曲线. (c) PBS和SubPc 4在4T1细胞中560~580 nm激光照射前后的Calcein-AM和PI双染色图像. 比例尺100 µm

Figure 7. (a) In vitro phototoxicity and dark toxicity of SubPc 4 against 4T1 cells evaluated by MTT assay. Cells were incubated with various concentrations of SubPc 4 (2.5, 5.0, 10.0, 20.0, 50.0 μg/mL) for 24 h, followed by irradiation with a 560~580 nm laser (0.5 W/cm2) for 10 min (phototoxicity group) or maintained in the dark (dark toxicity group). Data are presented as mean±SD (n=3). (b) Dose-response curve fitted from the phototoxicity data in (a) and the corresponding half‑maximal inhibitory concentration. (c) Calcein-AM and PI double-staining images of 4T1 cells treated with PBS and SubPc 4 before and after 560~580 nm laser irradiation. The scale bar is 100 µm

图8. SubPc 4对荷瘤小鼠的光动力治疗疗效及生物安全性评价分组: PBS＋Laser、SubPc 4、SubPc 4＋Laser. 光照条件: 560~580 nm 激光, 0.5 W/cm2, 照射10 min. (a)离体肿瘤图; (b)小鼠体重变化曲线; (c)离体肿瘤质量统计图; (d)肿瘤体积生长曲线; (e) 肿瘤组织H&E染色, 比例尺50 µm; (f)心、肝、脾、肺、肾H&E染色(数据为均值±SD), 比例尺50 µm

Figure 8. Photodynamic therapy efficacy and biosafety evaluation of SubPc 4 in tumor-bearing mice. Groups: PBS＋Laser, SubPc 4 alone, and SubPc 4 ＋Laser. Laser irradiation: 560~580 nm, 0.5 W/cm2, 10 min. (a) Representative photographs of excised tumors; (b) Body weight changes of mice; (c) Statistical comparison of excised tumor weights; (d) Tumor volume growth curves; (e) H&E staining of tumor tissues, The scale bar is 50 µm; (f) H&E staining of heart, liver, spleen, lung, and kidney (Data are shown as mean±SD), The scale bar is 50 µm

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