基于共轭大环的近红外超分子光敏剂用于乏氧光动力治疗

doi:10.6023/A26020061

摘要/Abstract

阳离子共轭大环具有较大的疏水空腔和良好的水溶性, 能够作为性能优异的主体与竹红菌素客体自组装成超分子光敏剂. 然而天然竹红菌素在光疗窗口的吸收波长短, 限制了超分子光敏剂在临床上的广泛应用. 在本工作中, 选用2位胺基取代的竹红菌素衍生物HCEA作为客体分子, 不仅在600~700 nm具有较强的宽吸收, 还可作为Ⅰ型光敏剂用于乏氧光动力治疗. 将HCEA分别与两种阳离子共轭大环AnBox^4＋和ExAnBox^4＋进行主客体组装得到了两种超分子光敏剂HCEA⊂AnBox^4＋和HCEA⊂ExAnBox^4＋, 不仅保留了HCEA的近红外特征吸收, 还增加了HCEA的水溶性, 主客体络合计量比为1∶1, 结合常数达到10⁶~10⁸数量级. 活性氧测试结果表明HCEA⊂AnBox^4＋和HCEA⊂ExAnBox^4＋均为Ⅰ型光敏剂, 在乏氧条件下具有优异的光动力治疗效果. 本研究为构建基于阳离子共轭大环的近红外超分子光敏剂和乏氧肿瘤的光动力治疗提供了新的思路.

关键词: 竹红菌素衍生物, 阳离子共轭大环, Ⅰ型光敏剂, 近红外超分子光敏剂, 乏氧光动力治疗

Cationic conjugated macrocycles, characterized by their large hydrophobic cavities and excellent water solubility, can serve as high-performance hosts to self-assemble with hypocrellin-based guests into supramolecular photosensitizers. However, the short absorption wavelength of supramolecular photosensitizers based on natural hypocrellins severely restricts their application in deep-tissue photodynamic therapy (PDT). In this work, HCEA was selected as the guest molecule which is a hypocrellin derivative with an amino substitution at the 2-position. HCEA exhibits broad-band absorption ranging from 600 to 700 nm and acts as a type-I photosensitizer, thereby offering enhanced potential for hypoxic photodynamic therapy. Through supramolecular assembly of HCEA with two water-soluble cationic conjugated macrocycles, namely AnBox^4＋ and ExAnBox^4＋, two supramolecular photosensitizers were successfully constructed, HCEA⊂AnBox^4＋ and HCEA⊂ExAnBox^4＋. UV-vis absorption spectroscopy confirmed that these host-guest complexes retained the near-infrared (NIR) absorption characteristics of HCEA and significantly improved the water solubility, addressing a long-standing challenge associated with hypocrellin-based photosensitizers. Fluorescence titration studies confirmed a 1∶1 host-guest stoichiometry between HCEA and the conjugated macrocycles, accompanied by remarkably high binding constants on the order of 10⁶ to 10⁸ L/mol, indicating strong and stable complex formation. Reactive oxygen species detection assays confirmed that both HCEA⊂AnBox^4＋and HCEA⊂ExAnBox^4＋ function as type-I supramolecular photosensitizers and can efficiently generate hydroxyl radicals (•OH) and superoxide anions ($\text{O}_{2}^{\bullet - }$). Confocal laser scanning microscopy further validated the efficient intracellular ROS generation by these supramolecular photosensitizers. Importantly, in vitro cytotoxicity assessments combined with calcein AM and propidium iodide (AM/PI) cell viability assays revealed that the supramolecular photosensitizers not only exhibited excellent biocompatibility but also achieved remarkable photodynamic therapeutic efficacy even under hypoxic conditions. This study provides a novel strategy for constructing conjugated macrocycle-based NIR supramolecular photosensitizers and implementing photodynamic therapy against hypoxic tumors, with promising potential for future clinical translation.

Key words: hypocrellin derivative, cationic conjugated macrocycle, type Ⅰ photosensitizer, near-infrared supramolecular photosensitizer, hypoxic photodynamic therapy

引用此文

苑玥珂, 董翔宇, 牧童, 高泽坤, 郑秀丽, 丛欢, 刘卫敏, 汪鹏飞. 基于共轭大环的近红外超分子光敏剂用于乏氧光动力治疗[J]. 化学学报, 2026, 84(6): 820-827.

Yueke Yuan, Xiangyu Dong, Tong Mu, Zekun Gao, Xiuli Zheng, Huan Cong, Weimin Liu, Pengfei Wang. Near-Infrared Supramolecular Photosensitizers Based on Conjugated Macrocycle for Hypoxic Photodynamic Therapy[J]. Acta Chimica Sinica, 2026, 84(6): 820-827.

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图/表 6

图1. HCEA, AnBox4＋, ExAnBox4＋以及1∶1主客体络合物的1H NMR谱(0.5 mmol/L, 400 MHz, CD3CN, 25 ℃)

Figure 1. 1H NMR spectra of HCEA, AnBox4＋, ExAnBox4＋ and 1∶1 host-guest complex (0.5 mmol/L, 400 MHz, CD3CN, 25 ℃)

图2. (a) HCEA (10 μmol/L)在乙腈和水溶液(ϕDMSO=20%)中的吸收和荧光光谱, 25 ℃; (b)纯水中等物质的量AnBox4＋和HCEA⊂AnBox4＋ (10 μmol/L)的归一化紫外-可见吸收光谱, 25 ℃; (c)纯水中等物质的量ExAnBox4＋和HCEA⊂ExAnBox4＋ (10 μmol/L)的归一化紫外-可见吸收光谱, 25 ℃

Figure 2. (a) UV-vis absorption and fluorescence spectra of HCEA (10 μmol/L) in CH3CN and H2O (ϕDMSO=20%) at 25 ℃; (b) Normalized UV-vis absorption spectra of equimolar AnBox4＋ and HCEA⊂AnBox4＋ (10 μmol/L) in H2O at 25 ℃; (c) Normalized UV-vis absorption spectra of equimolar ExAnBox4＋ and HCEA⊂ExAnBox4＋ (10 μmol/L) in H2O at 25 ℃

图3. (a) 在HCEA⊂AnBox4＋, HCEA⊂ExAnBox4＋, AnBox4＋或ExAnBox4＋存在下, DCFH在525 nm处荧光强度的相对变化(635 nm, 20 mW/cm2); (b)~(d)在MB, HCEA⊂AnBox4＋或HCEA⊂ExAnBox4＋存在下, ABDA吸收光谱的变化(635 nm, 5 mW/cm2) (pH 7.4 PBS, 635 nm处吸光度为0.1, 25 ℃)

Figure 3. (a) Relative changes in fluorescence intensity at 525 nm of DCFH within HCEA⊂AnBox4＋, HCEA⊂ExAnBox4＋, AnBox4＋ or ExAnBox4＋ (635 nm, 20 mW/cm2); (b)~(d) The changes of absorption spectra of ABDA within MB, HCEA⊂AnBox4＋ or HCEA⊂ExAnBox4＋ (635 nm, 5 mW/cm2) (pH 7.4 PBS, The absorbance at 635 nm is 0.1, 25 ℃)

图4. (a)在HCEA⊂AnBox4＋, HCEA⊂ExAnBox4＋, AnBox4＋或ExAnBox4＋存在下, HPF在526 nm处荧光强度的相对变化(635 nm, 50 mW/cm2); (b)在HCEA⊂AnBox4＋, HCEA⊂ExAnBox4＋, AnBox4＋或ExAnBox4＋存在下, DHR123在527 nm处荧光强度的变化(635 nm, 50 mW/cm2); (c) HCEA⊂AnBox4＋和HCEA⊂ExAnBox4＋在水中的稳定性测试(635 nm, 100 mW/cm2); (d)黑暗条件下HCEA⊂AnBox4＋和HCEA⊂ExAnBox4＋在pH7.4的PBS中稳定性测试

Figure 4. (a) Relative changes in fluorescence intensity at 526 nm of HPF within HCEA⊂AnBox4＋, HCEA⊂ExAnBox4＋, AnBox4＋or ExAnBox4＋ (635 nm, 50 mW/cm2); (b) The changes of fluorescence intensity of DHR123 at 527 nm within HCEA⊂AnBox4＋, HCEA⊂ExAnBox4＋, AnBox4＋or ExAnBox4＋(635 nm, 50 mW/cm2); (c) The stability of HCEA⊂AnBox4＋ and HCEA⊂ExAnBox4＋ detected via absorbance spectra (635 nm, 100 mW/cm2); (d) The stability of HCEA⊂AnBox4＋ and HCEA⊂ExAnBox4＋ detected in pH 7.4 PBS stored in dark

图5. (a)超分子光敏剂HCEA⊂AnBox4＋或HCEA⊂ExAnBox4＋分别与Hoechst共染后的荧光成像; (b)采用DCFH-DA检测HCEA⊂AnBox4＋或HCEA⊂ExAnBox4＋在HeLa细胞内常氧和乏氧条件下产生的ROS荧光成像; (c)采用DHE检测HCEA⊂AnBox4＋或HCEA⊂ExAnBox4＋在HeLa细胞内常氧和乏氧条件下产生的$\text{O}_{2}^{\bullet - }$荧光成像; (d)采用HPF检测HCEA⊂AnBox4＋或HCEA⊂ExAnBox4＋在HeLa细胞内常氧和乏氧条件下产生的•OH荧光成像. 标尺: 25 μm

Figure 5. (a) Fluorescence images of supramolecular photosensitizers HCEA⊂AnBox4＋ or HCEA⊂ExAnBox4＋ co-incubated with Hoechst; (b) Fluorescence images of ROS production detected by DCFH-DA in HeLa cells treated with HCEA⊂AnBox4＋ or HCEA⊂ExAnBox4＋ under normoxic or hypoxic conditions; (c) Fluorescence images of $\text{O}_{2}^{\bullet - }$ production detected by DHE in HeLa cells treated with HCEA⊂AnBox4＋ or HCEA⊂ExAnBox4＋ under normoxic or hypoxic conditions; (d) Fluorescence images of •OH production detected by HPF in HeLa cells treated with HCEA⊂AnBox4＋ or HCEA⊂ExAnBox4＋ under normoxic or hypoxic conditions. Scale bar: 25 μm

图6. HeLa细胞与(a) HCEA⊂AnBox4＋和(b) HCEA⊂ExAnBox4＋在常氧和乏氧条件下孵育2 h并光照后的细胞活力检测(635 nm, 100 mW/cm2); (c) HeLa细胞与HCEA⊂AnBox4＋或HCEA⊂ExAnBox4＋孵育2 h后的暗毒性; (d) HeLa细胞孵育不同浓度的HCEA⊂AnBox4＋或HCEA⊂ExAnBox4＋并光照后的AM/PI荧光成像实验(635 nm, 100 mW/cm2). 标尺: 100 μm

Figure 6. Relative viability of HeLa cells incubated 2 h with (a) HCEA⊂AnBox4＋and (b) HCEA⊂ExAnBox4＋ under both normoxic and hypoxic conditions after laser irradiation (635 nm, 100 mW/cm2); (c) Dark toxicity of HeLa cells incubated with HCEA⊂AnBox4＋ or HCEA⊂ExAnBox4＋; (d) Fluorescence images of the calcein AM and propidium iodide (PI) incubated with HCEA⊂AnBox4＋ or HCEA⊂ExAnBox4＋ after laser irradiation (635 nm, 100 mW/cm2). Scale bar: 100 μm

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