Near-Infrared Supramolecular Photosensitizers Based on Conjugated Macrocycle for Hypoxic Photodynamic Therapy

doi:10.6023/A26020061

Abstract

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

Cite this article

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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Fig. & Tab. 6

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

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 ℃)

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

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

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

References 41

[1]	Celli, J. P.; Spring, B. Q.; Rizvi, I.; Evans, C. L.; Samkoe, K. S.; Verma, S.; Pogue, B. W.; Hasan, T. Chem. Rev. 2010, 110, 2795. doi: 10.1021/cr900300p
[2]	Chinna Ayya Swamy, P.; Sivaraman, G.; Priyanka, R. N.; Raja, S. O.; Ponnuvel, K.; Shanmugpriya, J.; Gulyani, A. Coord. Chem. Rev. 2020, 411, 213233. doi: 10.1016/j.ccr.2020.213233
[3]	Kwiatkowski, S.; Knap, B.; Przystupski, D.; Saczko, J.; Kędzierska, E.; Knap-Czop, K.; Kotlińska, J.; Michel, O.; Kotowski, K.; Kulbacka, J. Biomed. Pharmacother. 2018, 106, 1098. doi: S0753-3322(18)34161-1 pmid: 30119176
[4]	Gomer, C. J.; Ferrario, A.; Luna, M.; Rucker, N.; Wong, S. Lasers Surg. Med. 2006, 38, 516. doi: 10.1002/lsm.v38:5
[5]	Singh, P. P.; Sinha, S.; Gahtori, P.; Mishra, D. N.; Pandey, G.; Srivastava, V. Dyes Pigm. 2024, 229, 112262. doi: 10.1016/j.dyepig.2024.112262
[6]	Yan, Y. H.; Liang, P. Z.; Zou, Y.; Yuan, L.; Peng, X. J.; Fan, J. L.; Zhang, X. B. Acta Chim. Sinica 2023, 81, 1642 (in Chinese). doi: 10.6023/A23050243
	(闫英红, 梁平兆, 邹杨, 袁林, 彭孝军, 樊江莉, 张晓兵, 化学学报, 2023, 81, 1642.) doi: 10.6023/A23050243
[7]	Li, Y.; Wang, W.; Zhang, Y. T.; Xiao, S. Z.; Lan, H.; Geng, P. Acta Chim. Sinica 2024, 82, 443 (in Chinese). doi: 10.6023/A24010035
	(李燕, 王玮, 张毓婷, 肖述章, 兰海闯, 耿鹏, 化学学报, 2024, 82, 443.) doi: 10.6023/A24010035
[8]	Huang, Y. Q.; Luo, J. W.; Li, J. Q.; Zhang, R.; Liu, X. F.; Fan, Q. L.; Huang, W. Acta Chim. Sinica 2024, 82, 903 (in Chinese). doi: 10.6023/A24040115
	(黄艳琴, 罗集文, 李佳启, 张瑞, 刘兴奋, 范曲立, 黄维, 化学学报, 2024, 82, 903.) doi: 10.6023/A24040115
[9]	Li, J. Q.; Huang, Y. Q.; Ding, G. C.; Zhang, R.; Liu, X. F.; Fan, Q. L.; Huang, W. Acta Chim. Sinica 2025, 83, 1324 (in Chinese). doi: 10.6023/A25050141
	(李佳启, 黄艳琴, 丁桂策, 张瑞, 刘兴奋, 范曲立, 黄维, 化学学报, 2025, 83, 1324.) doi: 10.6023/A25050141
[10]	Yu, Q. S.; Qi, C.; Gu, S. X.; Yang, X. D.; Jiang, Q.; Wei, R. B.; Shi, P. F. Acta Chim. Sinica 2025, 83, 579 (in Chinese). doi: 10.6023/A24120380
	(于千水, 戚聪, 顾顺心, 杨欣达, 姜秦, 魏荣斌, 施鹏飞, 化学学报, 2025, 83, 579.) doi: 10.6023/A24120380
[11]	Aziz, B.; Aziz, I.; Khurshid, A.; Raoufi, E.; Esfahani, F. N.; Jalilian, Z.; Mozafari, M. R.; Taghavi, E.; Ikram, M. Biomedicines 2023, 11, 224. doi: 10.3390/biomedicines11010224
[12]	Mohammadi, A.; Mansoori, B.; Baradaran, B. Biomed. Pharmacother. 2017, 86, 262. doi: S0753-3322(16)31747-4 pmid: 28006752
[13]	Mansoori, B.; Mohammadi, A.; Amin Doustvandi, M.; Mohammadnejad, F.; Kamari, F.; Gjerstorff, M. F.; Baradaran, B.; Hamblin, M. R. Photodiagn. Photodyn. Ther. 2019, 26, 395. doi: 10.1016/j.pdpdt.2019.04.033
[14]	Zhou, X. X.; Ying, X. F.; Wu, L. J.; Liu, L. Q.; Wang, Y.; He, Y.; Han, M. Planta Med. 2024, 90, 368. doi: 10.1055/a-2257-9194
[15]	Zhang, Y.; Xie, J.; Zhang, L. Y.; Li, C.; Chen, H. X.; Gu, Y.; Zhao, J. Q. Photochem. Photobiol. Sci. 2009, 8, 1676. doi: 10.1039/b9pp00046a
[16]	Li, X. C.; Liu, Y. L.; Wu, L. L.; Zhao, J. ChemMedChem 2024, 19, e202300599. doi: 10.1002/cmdc.v19.4
[17]	Wu, J. S.; Sha, J.; Zhang, C. L.; Liu, W. M.; Zheng, X. L.; Wang, P. F. VIEW 2020, 1, 20200090. doi: 10.1002/viw2.v1.3
[18]	Lan, M. H.; Zhao, S. J.; Liu, W. M.; Lee, C. S.; Zhang, W. J.; Wang, P. F. Adv. Healthcare Mater. 2019, 8, 1900132. doi: 10.1002/adhm.v8.13
[19]	Zhao, X. Z.; Liu, J. P.; Fan, J. L.; Chao, H.; Peng, X. J. Chem. Soc. Rev. 2021, 50, 4185. doi: 10.1039/D0CS00173B
[20]	Zhang, Y. Q.; Zhan, C.; Gong, Y.; Lu, F.; Fan, Q. L. Acta Chim. Sinica 2024, 82, 1226 (in Chinese). doi: 10.6023/A24090286
	(张雨琦, 占辰, 贡祎, 陆峰, 范曲立, 化学学报, 2024, 82, 1226.) doi: 10.6023/A24090286
[21]	Li, X. S.; Lee, S. Y.; Yoon, J. Y. Chem. Soc. Rev. 2018, 47, 1174. doi: 10.1039/C7CS00594F
[22]	Zhang, T.-X.; Zhang, Z.-Z.; Yue, Y.-X.; Hu, X.-Y.; Huang, F.; Shi, L.; Liu, Y.; Guo, D.-S. Adv. Mater. 2020, 32, 1908435. doi: 10.1002/adma.v32.28
[23]	Liu, K.; Liu, Y. L.; Yao, Y. X.; Yuan, H. X.; Wang, S.; Wang, Z. Q.; Zhang, X. Angew. Chem., Int. Ed. 2013, 52, 8285. doi: 10.1002/anie.v52.32
[24]	Liu, Y.-Y.; Yu, X.-Y.; Pan, Y.-C.; Yin, H.; Chao, S.; Li, Y.; Ma, H.; Zuo, M.; Teng, K.-X.; Hou, J.-L.; Chen, Y.; Guo, D.-S.; Wang, R.; Pei, Y.; Pei, Z.; Xu, J.-F.; Hu, X.-Y.; Li, C.; Yang, Q.-Z.; Wang, L.; Liu, Y.; Li, Z.-T. Sci. China Chem. 2024, 67, 1397. doi: 10.1007/s11426-024-1971-4
[25]	Liao, Z.-H.; Wang, F. Smart Mol. 2024, 2, e20240036.
[26]	Roy, I.; Bobbala, S.; Young, R. M.; Beldjoudi, Y.; Nguyen, M. T.; Cetin, M. M.; Cooper, J. A.; Allen, S.; Anamimoghadam, O.; Scott, E. A.; Wasielewski, M. R.; Stoddart, J. F. J. Am. Chem. Soc. 2019, 141, 12296. doi: 10.1021/jacs.9b03990
[27]	Xing, C. Y.; Chen, H. K.; Guan, Y. P.; Zhang, S. Q.; Tong, T. Y.; Ding, N.; Luo, T. T.; Kang, Y.; Pang, J. J. Colloid Interface Sci. 2022, 628, 864. doi: 10.1016/j.jcis.2022.08.110
[28]	Schmitt, F.; Freudenreich, J.; Barry, N. P. E.; Juillerat-Jeanneret, L.; Süss-Fink, G.; Therrien, B. J. Am. Chem. Soc. 2012, 134, 754. doi: 10.1021/ja207784t
[29]	Peng, H.-Q.; Niu, L.-Y.; Chen, Y.-Z.; Wu, L.-Z.; Tung, C.-H.; Yang, Q.-Z. Chem. Rev. 2015, 115, 7502. doi: 10.1021/cr5007057
[30]	Fan, Y. Q.; He, J.; Guo, S. Z.; Jiang, H. ChemPlusChem 2024, 89, e202300536. doi: 10.1002/cplu.v89.7
[31]	Kumar, P.; Gupta, D.; Grewal, S.; Srivastava, A.; Kumar Gaur, A.; Venkataramani, S. Chem. Rec. 2022, 22, e202200074. doi: 10.1002/tcr.v22.11
[32]	Krieg, E.; Bastings, M. M. C.; Besenius, P.; Rybtchinski, B. Chem. Rev. 2016, 116, 2414. doi: 10.1021/acs.chemrev.5b00369
[33]	Roy, R.; Brouillac, C.; Jacques, E.; Quinton, C.; Poriel, C. Angew. Chem., Int. Ed. 2024, 63, e202402608. doi: 10.1002/anie.v63.30
[34]	Zheng, X. L.; Lei, S.-N.; Gao, Z. K.; Dong, X. Y.; Xiao, H. Y.; Liu, W. M.; Tung, C.-H.; Wu, L.-Z.; Wang, P. F.; Cong, H. Chem. Sci. 2023, 14, 3523. doi: 10.1039/D3SC00107E
[35]	Gao, Z. K.; Zheng, X. L.; Dong, X. Y.; Liu, W. M.; Sha, J.; Bian, S. S.; Li, J.; Cong, H.; Lee, C.; Wang, P. Adv. Healthcare Mater. 2024, 13, 2401778. doi: 10.1002/adhm.v13.28
[36]	Mu, T.; Zheng, X. L.; Yuan, Y. K.; Wang, Y. P.; Liu, T.; Li, H. X.; Wu, J. S.; Li, F.; Zhang, W. J.; Lee, C.-S.; Liu, W. M.; Wang, P. F. Sci. China Mater. 2025, 68, 4575. doi: 10.1007/s40843-025-3548-9
[37]	Tian, J. W.; Li, B. W.; Zhang, F.; Yao, Z.; Song, W. T.; Tang, Y. F.; Ping, Y.; Liu, B. Angew. Chem., Int. Ed. 2023, 62, e202307288. doi: 10.1002/anie.v62.46
[38]	Huang, L.; Zhao, S. J.; Wu, J.; Yu, L.; Singh, N.; Yang, K.; Lan, M. H.; Wang, P. F.; Kim, J. S. Coord. Chem. Rev. 2021, 438, 213888. doi: 10.1016/j.ccr.2021.213888
[39]	Yu, L.; Liu, Z.; Xu, W.; Jin, K.; Liu, J. L.; Zhu, X. H.; Zhang, Y.; Wu, Y. H. Acta Pharm. Sin. B 2024, 14, 1111. doi: 10.1016/j.apsb.2023.11.007
[40]	Chen, D. P.; Xu, Q.; Wang, W. J.; Shao, J. J.; Huang, W.; Dong, X. C. Small 2021, 17, 2006742. doi: 10.1002/smll.v17.31
[41]	Brynn Hibbert, D.; Thordarson, P. Chem. Commun. 2016, 52, 12792. doi: 10.1039/C6CC03888C

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