A Catalyst-Free Example of Photomediated Biological Orthogonal Fluorescence Probes

doi:10.6023/A25040102

Acta Chimica Sinica ›› 2025, Vol. 83 ›› Issue (8): 816-826.DOI: 10.6023/A25040102 Previous Articles Next Articles

Article

无需催化剂的光介导生物正交荧光探针实例

赵玉强^a, 那迪^a, 和晓波^a, 朱立平^b, 周莹^b^,^*(), 曾广智^a^,^*(), 樊保敏^a^,^*()

^a 云南民族大学化学与环境学院、云南省手性功能物质研究与利用重点实验室昆明 650504
^b 云南大学化学科学与工程学院昆明 650091

投稿日期:2025-04-01 发布日期:2025-05-08
通讯作者: 周莹, 曾广智, 樊保敏
基金资助:
云南省手性功能物质研究与利用重点实验室(202402AN360010); 云南省科技厅-基础研究专项-重大项目(202401BC070018); 云南省科技厅-云南大学联合特殊项目(202201BF070001-001); 国家自然科学基金(22067019); 国家自然科学基金(22367023)

A Catalyst-Free Example of Photomediated Biological Orthogonal Fluorescence Probes

Yu-Qiang Zhao^a, Di Na^a, Xiaobo He^a, Liping Zhu^b, Ying Zhou^b^,^*(), Guang-Zhi Zeng^a^,^*(), Baomin Fan^a^,^*()

^a Yunnan Key Laboratory of Chiral Functional Substance Research and Application, School of Chemistry & Environment, Yunnan Minzu University, Kunming 650504, Yunnan, China
^b College of Chemical Science and Technology, Yunnan University, Kunming 650091, Yunnan, China

Received:2025-04-01 Published:2025-05-08
Contact: Ying Zhou, Guang-Zhi Zeng, Baomin Fan
Supported by:
Yunnan Key Laboratory of Chiral Functional Substance Research and Application(202402AN360010); Yunnan Provincial Department of Science and Technology-Basic Research-Major project(202401BC070018); Yunnan Provincial Science and Technology Department-Yunnan University Joint Special Project(202201BF070001-001); National Natural Science Foundation of China(22067019); National Natural Science Foundation of China(22367023)

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Abstract

A bioorthogonal fluorescent probe, NATz, was meticulously crafted by incorporating 1,8-naphthalimide onto a 1,2,4,5-tetrazine group endowed with bioorthogonal reaction activity. The spectral examination revealed the activation of fluorescence following the interaction between NATz and trans-cyclooctene (TCO) in a phosphate buffer saline (PBS) solution. Systematic time-dependent density functional theory (TDDFT) calculations elucidate that the fluorescence quenching and activation of NATz are predominantly mediated by a photoinduced electron transfer (PET) mechanism, involving excited-state charge redistribution between the tetrazine moiety and naphthalimide fluorophore. Significantly, the UV light at 365 nm substantially accelerated the reaction rate between NATz and TCO. High-resolution mass spectrometry (HRMS) confirmed that 365 nm UV irradiation exclusively promotes inverse electron-demand Diels-Alder (IEDDA) adduct formation, with no detectable photodegradation byproducts under the experimental conditions. Integrated with reaction pathway calculations, these data conclusively establish that light enhances the bioorthogonal IEDDA reaction between 1,2,4,5-tetrazine (NATz) and TCO by photon energy transfer surpassing the reaction energy barrier. The MTS cytotoxicity assay revealed that NATz, even at elevated concentrations (40 μmol/L), maintained cell proliferation inhibition rates below 30% across five cell lines (LO2, BEAS-2B, HeLa, HepG2, and A549), thereby confirming its favorable biocompatibility. Laser scanning confocal microscopy (LSCM) confirms NAtz's suitability for light-activated bioimaging applications, achieving cellular-resolution visualization in single cells and intact multicellular organisms (e.g., Caenorhabditis elegans). In order to further verify the applicability of light-mediated bioorthogonal fluorescent probes, a lysosome-targeted morpholine group was introduced on the basis of NATz, and the lysosome-targeted bioorthogonal fluorescent probe Lyso-NATz was prepared. The spectral test results indicated that after the introduction of the morpholine group, this series of probes still maintained the characteristics of photomediated bioorthogonal imaging. Meanwhile, the cytotoxicity experiment proved that the introduction of the morpholine group did not affect its biocompatibility. Finally, Lyso-NATz was used to achieve photomediated bioorthogonal fluorescence imaging targeting lysosomes at the cellular level.

Key words: photomediated, bioorthogonal chemistry, fluorescence probe, tetrazine, targeting to lysosomes

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Yu-Qiang Zhao, Di Na, Xiaobo He, Liping Zhu, Ying Zhou, Guang-Zhi Zeng, Baomin Fan. A Catalyst-Free Example of Photomediated Biological Orthogonal Fluorescence Probes[J]. Acta Chimica Sinica, 2025, 83(8): 816-826.

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Fig. & Tab. 8

Scheme 1. Schematic representation of the synthesis route of NATz

Figure 1. (a), (c) The measured (solid line) and calculated (dashed line) UV spectra; The hole charge distribution and frontier orbital contributions for the fourth excited state of NATz (b) and NATz-TCO (d) with blue isosurfaces representing holes and green isosurfaces representing charges. (e) The fluorescence spectrum (solid line) and excitation spectrum (dashed line) of NATz after the reaction with TCO. (f) The fluorescence intensity of NATz after reacting with TCO in different solvents (H->L represents the transition from the highest occupied molecular orbital (HOMO) to the lowest unoccupied molecular orbital (LUMO))

Figure 2. (a) Conceptual illustration predicting the PET process through the energy level difference of frontier orbitals; (b) Schematic diagram depicting the orbital energy level difference and electron distribution of NATz; (c) Depiction of NATz's excited state information and corresponding distributions of frontier orbital electrons; (d) Excited state information of NATz-TCO along with corresponding distributions of frontier orbital electrons

Figure 3. (a)~(c) Fluorescence spectra of NATz under diverse conditions; (d) Fluorescence intensity at 450 nm under diverse conditions. 1: NATz, 2: NATz＋TCO (1 h), 3: NATz＋TCO (24 h), 4: NATz＋TCO (48 h), 5: NATz＋TCO (72 h), 6: NATz＋TCO (1 h)＋UV (10 min), 7: NATz＋UV (10 min)

Figure 4. (a) Conceptual diagram depicting the photocatalytic reaction between TCO and tetrazine; (b) High-resolution mass spectrum of NATz; (c) High-resolution mass spectrum of NATz after one hour of reaction with TCO; (d) High-resolution mass spectrum of NATz after one hour of reaction with TCO followed by 10 min of UV light exposure

Figure 5. (a) Illustration of the IEDDA reaction pathway between tetrazine and TCO; (b) Transition states and reaction pathway for the cycloaddition reaction between tetrazine and TCO; (c) Energy profiles of different intermediates in the cycloaddition process of tetrazine and TCO

Figure 6. (a) Photocatalytic orthogonal imaging of NATz within HepG2 cells; (b) Photocatalytic orthogonal imaging of NATz inside the body of elegant hidden worms (C. elegans); (c) Cellular toxicity assessment of NATz (40 μmol/L); (d) Average fluorescence intensity within cells as depicted in (a); (e) Average fluorescence intensity within worms as shown in (d). scale bar: 10 μm/200 μm. **: P<0.01, ***: P<0.001

Figure 7. (a) Synthesis pathway of Lyso-NATz; (b) Photocatalytic orthogonal fluorescence spectra of Lyso-NATz; (c) Evaluation of cellular toxicity for Lyso-NATz; (d) Co-localization imaging of Lyso-NATz with lysosomal markers (LysoTracker Red). scale bar: 10 μm

References 52

[1]	Liao, Y. M.; Wu, B. Q.; Tang, R. Z. Prog. Chem. 2022, 34, 2134 (in Chinese).
	(廖伊铭, 吴宝琪, 唐荣志, 化学进展, 2022, 34, 2134.) doi: 10.7536/PC220103
[2]	Debets, M. F.; van Hest, J. C. M.; Rutjes, F. P. J. T. Org. Biomol. Chem. 2013, 11, 6439.
[3]	Schoch, J.; Ameta, S.; Jäschke, A. Chem. Commun. 2011, 47, 12536.
[4]	Javaherian, M.; Kazemi, F.; Ghaemi, M. Chin. Chem. Lett. 2014, 25, 1643.
[5]	Zhang, Q.; Huang, W. J.; Wang, Y. B.; Li, X. J.; Zhang, Y. H. Prog. Chem. 2020, 32, 147 (in Chinese).
	(章强, 黄文峻, 王延斌, 李兴建, 张宜恒, 化学进展, 2020, 32, 147.) doi: 10.7536/PC190804
[6]	Wang, C.; Zhou, F.; Zhou, J. Chin. J. Org. Chem. 2020, 40, 3065 (in Chinese).
	(王才, 周锋, 周剑, 有机化学, 2020, 40, 3065.) doi: 10.6023/cjoc202005020
[7]	Weterings, J.; Rijcken, C. J. F.; Veldhuis, H.; Meulemans, T.; Hadavi, D.; Timmers, M.; Honing, M.; Ippeld, H.; Liskamp, R. M. J. Chem. Sci. 2020, 11, 9011. doi: 10.1039/d0sc03477k pmid: 34123155
[8]	Agard, N. J.; Prescher, J. A.; Bertozzi, C. R. J. Am. Chem. Soc. 2005, 127, 11196.
[9]	Li, X. R.; Fang, T.; Boons, G. J. Angew. Chem. Int. Ed. 2014, 53, 7179.
[10]	Zhou, Y. C.; Zhou, Z.; Du, W.; Chen, Y. C. Acta Chim. Sinica 2018, 76, 382 (in Chinese).
	(周远春, 周志, 杜玮, 陈应春, 化学学报, 2018, 76, 382.) doi: 10.6023/A18040131
[11]	Lv, J.; Zhong, X. R.; Cheng, J. P.; Luo, S. Z. Acta Chim. Sinica 2012, 70, 1518 (in Chinese).
	(吕健, 钟兴仁, 程津培, 罗三中, 化学学报, 2012, 70, 1518.) doi: 10.6023/A12060346
[12]	Zhang, X. R.; Luo, H. X.; Fan, X. Y.; Chen, P. Sci. Sin. Chim. 2020, 50, 1280 (in Chinese).
	(张贤睿, 罗惠鑫, 樊新元, 陈鹏, 中国科学: 化学, 2020, 50, 1280.)
[13]	Oliveira, B. L.; Guo, Z.; Bernardes, G. J. L. Chem. Soc. Rev. 2017, 46, 4895. doi: 10.1039/c7cs00184c pmid: 28660957
[14]	Wen, X. D.; Zeng, W. H.; Zhang, J. Y.; Liu, Y. L.; Miao, Y. X.; Liu, S. H.; Yang, Y. L.; Xu, J. J.; Ye, D. J. Angew. Chem. Int. Ed. 2024, 63, e202314039.
[15]	Fang, G. M.; Wang, C.; Shi, J.; Guo, Q. X. Acta Chim. Sinica 2009, 67, 2335 (in Chinese).
	(方葛敏, 王晨, 石景, 郭庆祥, 化学学报, 2009, 67, 2335.)
[16]	Zhang, Y. Z.; Zhu, L. J.; Shao, X. L.; Xu, W. T. Biol. Bull. 2019, 35, 187 (in Chinese)
	(张洋子, 朱龙佼, 邵向丽, 许文涛, 生物技术通报, 2019, 35, 187.) doi: 10.13560/j.cnki.biotech.bull.1985.2018-0096
[17]	Bednarek, C.; Wehl, I.; Jung, N.; Schepers, U.; Bräse, S. Chem. Rev. 2020, 120, 4301.
[18]	Yang, M. Y.; Chen, P. Acta Chim. Sinica 2015, 73, 783 (in Chinese).
	(杨麦云, 陈鹏, 化学学报, 2015, 73, 783.) doi: 10.6023/A15030214
[19]	Wang, X.; Zhang, X. R.; Huang, Z. Y.; Fan, X. Y.; Chen, P. Acta Chim. Sinica 2021, 79, 406 (in Chinese). doi: 10.6023/A20110530
	(汪欣, 张贤睿, 黄宗煜, 樊新元, 陈鹏, 化学学报, 2021, 79, 406.) doi: 10.6023/A20110530
[20]	Yang, J.; Zhang, J.; Qi, L.; Hu, C.; Chen, Y. Chem. Commun. 2015, 51, 5275.
[21]	Huang, H.; Zhang, G.; Gong, L.; Zhang, S.; Chen, Y. J. Am. Chem. Soc. 2014, 136, 2280.
[22]	Li, J.; Kong, H.; Huang, L.; Cheng, B.; Qin, K.; Zheng, M.; Yan, Z.; Zhang, Y. J. Am. Chem. Soc. 2018, 140, 14542.
[23]	Zhang, L.; Zhang, X.; Yao, Z.; Jiang, S.; Deng, J.; Li, B.; Yu, Z. J. Am. Chem. Soc. 2018, 140, 7390.
[24]	Liang, X.; Qian, S.; Lou, Z.; Hu, R.; Hou, Y.; Chen, P. R.; Fan, X. Angew. Chem. Int. Ed. 2023, 62, e202310920.
[25]	Su, D. Y.; Li, J.; Pan, L. L.; Wu, H. X.; Mao, W. Y. Acta Pharm. Sin. 2021, 56, 1086 (in Chinese).
	(粟敦妍, 李杰, 潘立立, 吴昊星, 毛梧宇, 药学学报, 2021, 56, 1086.)
[26]	Wang, X. M.; Li, J.; Shen, G. H.; Pan, L. L.; Tian, R.; Sun, H. B.; Wu, H. X. Acta Pharm. Sin. 2020, 55, 1634 (in Chinese).
	(王晓蒙, 李杰, 沈国华, 潘立立, 田蓉, 孙洪宝, 吴昊星, 药学学报, 2020, 55, 1634.)
[27]	Mboyi, C. D.; Vivier, D.; Daher, A.; Fleurat-Lessard, P.; Cattey, H.; Devillers, C. H.; Bernhard, C.; Denat, F.; Roger, J.; Hierso, J. C. Angew. Chem. Int. Ed. 2020, 59, 1149.
[28]	Jemas, A.; Xie, Y. X.; Pigga, J. E.; Caplan, J. L.; Ende, C. W. A.; Fox, J. M. J. Am. Chem. Soc. 2022, 144, 1647.
[29]	Rosenberger, J. E.; Xie, Y. X.; Fang, Y. Z.; Lyu, X. Y.; Trout, W. S.; Dmitrenko, O.; Fox, J. M. J. Am. Chem. Soc. 2023, 145, 6067. doi: 10.1021/jacs.2c10655 pmid: 36881718
[30]	Zhang, H.; Trout, W. S.; Liu, S.; Andrade, G. A.; Hudson, D. A.; Scinto, S. L.; Dicker, K. T.; Li, Y.; Lazouski, N.; Rosenthal, J. J. Am. Chem. Soc. 2016, 138, 5978. doi: 10.1021/jacs.6b02168 pmid: 27078610
[31]	Wang, C. Q.; Zhang, H.; Zhang, T.; Zou, X. Y.; Wang, H.; Rosenberger, J. E.; Vannam, R.; Trout, W. S.; Grimm, J. B.; Lavis, L. D.; Thorpe, C.; Jia, X. Q.; Li, Z. B.; Fox, J. M. J. Am. Chem. Soc. 2021, 143, 10793.
[32]	Liu, L. P.; Zhang, D. Y.; Johnson, M.; Devaraj, N. K. Nat. Chem. 2022, 14, 1078.
[33]	Zheng, J. J.; Zhao, Y.; Truhlar, D. G. J. Chem. Theory Comput. 2009, 5, 808.
[34]	Strych, S.; Journot, G.; Pemberton, R. P.; Wang, S. C.; Tantillo, D. J. Angew. Chem. Int. Ed. 2015, 54, 5079.
[35]	Liu, Z. Y.; Lu, T.; Chen, Q. X. Carbon 2020, 165, 461.
[36]	Wu, H. X.; Devaraj, N. K. Acc. Chem. Res. 2018, 51, 1249.
[37]	Zhang, R. S.; Gao, J. K.; Zhao, G. X.; Zhou, L. M.; Kong, F. D.; Jiang, T.; Jiang, H. F. Nanoscale 2023, 15, 461.
[38]	van Onzen, A. H. A. M.; Versteegen, R. M.; Hoeben, F. J. M.; Filot, I. A. W.; Rossin, R.; Zhu, T.; Wu, J.; Hudson, P. J.; Janssen, H. M.; ten Hoeve, W.; Robillard, M. S. J. Am. Chem. Soc. 2020, 142, 10955.
[39]	Cao, W. B.; Wang, H. Y.; Quan, M.; Li, Y. X.; Su, Y. Y.; Li, Y. H.; Jiang, W.; Liu, T. Chem 2023, 9, 2881.
[40]	Fan, X. Y.; Ge, Y.; Lin, F.; Yang, Y.; Zhang, G.; Ngai, W. S. C.; Lin, Z.; Zheng, S. Q.; Wang, J.; Zhao, J. Y.; Li, J.; Chen, P. R. Angew. Chem. Int. Ed. 2016, 55, 14046.
[41]	Li, S. P. Y.; Yip, A. M. H.; Liu, H. W.; Lo, K. K. W. Biomaterials 2016, 103, 305.
[42]	Devaraj, N. K.; Hilderbrand, S.; Upadhyay, R.; Mazitschek, R.; Weissleder, R. Angew. Chem. Int. Ed. 2010, 49, 2869. doi: 10.1002/anie.200906120 pmid: 20306505
[43]	Carlson, J. C. T.; Meimetis, L. G.; Hilderbrand, S. A.; Weissleder, R. Angew. Chem. Int. Ed. 2013, 52, 6917.
[44]	Mao, W. Y.; Chi, W. J.; He, X. Y.; Wang, C.; Wang, X. Y.; Yang, H. J.; Liu, X. G.; Wu, H. X. Angew. Chem. Int. Ed. 2022, 61, e202117386.
[45]	Chi, W. J.; Chen, J.; Liu, W. J.; Wang, C.; Qi, Q. K.; Qiao, Q. L.; Tan, T. M.; Xiong, K. M.; Li, X.; Kang, K.; Chang, Y. T.; Xu, Z. C.; Liu, X. G. J. Am. Chem. Soc. 2020, 142, 6777.
[46]	Loredo, A.; Tang, J.; Wang, L. S.; Wu, K. L.; Peng, Z.; Xiao, H. Chem. Sci. 2020, 11, 4410. doi: 10.1039/d0sc01009j pmid: 33384859
[47]	Xu, M. M.; Cai, Q. Chin. J. Org. Chem. 2022, 42, 698 (in Chinese).
	(徐萌萌, 蔡泉, 有机化学, 2022, 42, 698.) doi: 10.6023/cjoc202109025
[48]	Zhu, Y. C.; Lin, F.; Liu, Z. Q.; Wang, X.; Wang, S. B.; Fan, X. Y.; Chen, P. R. Chin. J. Chem. 2025, 43, 553.
[49]	Yang, J.; Dai, J.; Lou, X.; Xia, F. Chin. Sci. Bull. 2020, 65, 3497 (in Chinese).
	(杨聚梁, 戴俊, 娄筱叮, 夏帆, 科学通报, 2020, 65, 3497.)
[50]	Zhang, Z. Q.; Yue, P. F.; Lu, T. Q.; Wang, Y.; Wei, Y. Q.; Wei, X. W. J. Hematol Oncol. 2021, 14, 79.
[51]	Frisch, M. J.; Trucks, G. W.; Schlegel, H. B.; Scuseria, G. E.; Robb, M. A.; Cheeseman, J. R.; Scalmani, G.; Barone, V.; Mennucci, B.; Petersson, G. A.; Nakatsuji, H.; Caricato, M.; Li, X.; Hratchian, H. P.; Izmaylov, A. F.; Bloino, J.; Zheng, G.; Sonnenberg, J. L.; Hada, M.; Ehara, M.; Toyota, K.; Fukuda, R.; Hasegawa, J.; Ishida, M.; Nakajima, T.; Honda, Y.; Kitao, O.; Nakai, H.; Vreven, T.; Montgomery, J. A., Jr.; Peralta, J. E.; Ogliaro, F.; Bearpark, M.; Heyd, J. J.; Brothers, E.; Kudin, K. N.; Staroverov, V. N.; Kobayashi, R.; Normand, J.; Raghavachari, K.; Rendell, A.; Burant, J. C.; Iyengar, S. S.; Tomasi, J.; Cossi, M.; Rega, N.; Millam, J. M.; Klene, M.; Knox, J. E.; Cross, J. B.; Bakken, V.; Adamo, C.; Jaramillo, J.; Gomperts, R.; Stratmann, R. E.; Yazyev, O.; Austin, A. J.; Cammi, R.; Pomelli, C.; Ochterski, J. W.; Martin, R. L.; Morokuma, K.; Zakrzewski, V. G.; Voth, G. A.; Salvador, P.; Dannenberg, J. J.; Dapprich, S.; Daniels, A. D.; Farkas, Ö.; Foresman, J. B.; Ortiz, J. V.; Cioslowski, J.; Fox, D. J. Gaussian 09, Revision E. 01, Gaussian, Inc., Wallingford CT, 2013.
[52]	Lu, T.; Chen, F. W. J. Comput. Chem. 2012, 33, 580.

无需催化剂的光介导生物正交荧光探针实例

A Catalyst-Free Example of Photomediated Biological Orthogonal Fluorescence Probes

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