Advances in Nitroreductase-Activated Fluorescent Probes Based on Photophysical Response Mechanisms

Chenye Jin; Jinmao You

doi:10.62177/jaet.v3i2.1392

Authors

Chenye Jin Shaoxing University
Jinmao You Shaoxing University

DOI:

https://doi.org/10.62177/jaet.v3i2.1392

Keywords:

Nitroreductase, Fluorescent Probe, Photoinduced Electron Transfer, Intramolecular Charge Transfer, Förster Resonance Energy Transfer, Hypoxia

Abstract

Nitroreductase (NTR) is a flavin mononucleotide-dependent oxidoreductase that selectively reduces aromatic nitro compounds to hydroxylamine or amino derivatives in the presence of NAD(P)H. Tumor hypoxia is often accompanied by upregulation of NTR levels, making it an ideal biomarker for hypoxia assessment. This article briefly reviews recent advances in NTR-activated fluorescent probes based on photophysical response mechanisms. According to their design principles, these probes are mainly classified into three categories: probes based on photoinduced electron transfer (PET), which utilize the nitro group to quench fluorescence and achieve a turn-on signal upon reduction; probes based on intramolecular charge transfer (ICT), which incorporate the nitro group into a donor-acceptor electronic system and produce spectral responses upon changes in electronic properties following reduction; and ratiometric probes based on Förster resonance energy transfer (FRET), which employ an internal reference channel to eliminate environmental interference. In addition, reversibly binding probes that block electron transfer by forming hydrogen bonds with the reduced cofactor represent a new mode of non-reactive detection. Finally, this review summarizes the core challenges faced by current probes and outlines future directions.

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References

Sharma, A., Arambula, J. F., Koo, S., Kumar, R., Singh, H., Sessler, J. L., & Kim, J. S. (2019). Hypoxia-targeted drug delivery. Chemical Society Reviews, 48(3), 771-813. https://doi.org/10.1039/C8CS00304A

Chen, Y., & Hu, L. (2009). Design of anticancer prodrugs for reductive activation. Medicinal Research Reviews, 29(1), 29-64. https://doi.org/10.1002/med.20137

Emami Nejad, A., Najafgholian, S., Rostami, A., Sistani, A., Shojaeifar, S., Esparvarinha, M., ... & Manian, M. (2021). The role of hypoxia in the tumor microenvironment and development of cancer stem cell: a novel approach to developing treatment. Cancer Cell International, 21(1), 62. https://doi.org/10.1186/s12935-020-01719-5

Vaupel, P. (2009). Prognostic potential of the pretherapeutic tumor oxygenation status. In Oxygen Transport to Tissue XXX (pp. 241-246). Boston, MA: Springer US.

Muz, B., De La Puente, P., Azab, F., & Kareem Azab, A. (2015). The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy. Hypoxia, 83-92. https://doi.org/10.2147/HP.S93413

Singleton, D. C., Macann, A., & Wilson, W. R. (2021). Therapeutic targeting of the hypoxic tumour microenvironment. Nature reviews Clinical oncology, 18(12), 751-772. https://doi.org/10.1038/s41571-021-00539-4

Bryant, D. W., McCalla, D. R., Leeksma, M., & Laneuville, P. (1981). Type I nitroreductases of Escherichia coli. Canadian journal of microbiology, 27(1), 81-86. https://doi.org/10.1139/m81-013

Wardman, P. (1985). Some reactions and properties of nitro radical-anions important in biology and medicine. Environmental Health Perspectives, 64, 309.

Peterson, F. J., Mason, R. P., Hovsepian, J., & Holtzman, J. L. (1979). Oxygen-sensitive and-insensitive nitroreduction by Escherichia coli and rat hepatic microsomes. Journal of Biological Chemistry, 254(10), 4009-4014.

Chen, Z., Han, F., Du, Y., Shi, H., & Zhou, W. (2023). Hypoxic microenvironment in cancer: molecular mechanisms and therapeutic interventions. Signal transduction and targeted therapy, 8(1), 70. https://doi.org/10.1038/s41392-023-01332-8

Cheng, D., Xu, W., Gong, X., Yuan, L., & Zhang, X. B. (2020). Design strategy of fluorescent probes for live drug-induced acute liver injury imaging. Accounts of Chemical Research, 54(2), 403-415. https://doi.org/10.1021/acs.accounts.0c00646

Xie, H., Li, Q., Yang, H., Gao, W., Zhang, Q., Zhang, P., & Ding, C. (2023). Mitochondrial-targeting fluorescent probe awakened by nitroreductase for hypoxic imaging and surgical navigation of solid tumors. Sensors and Actuators B: Chemical, 396, 134556. https://doi.org/10.1016/j.snb.2023.134556

Wickramasinghe, N. I., Corbin, B., Kanakarathna, D. Y., Pang, Y., Abeywickrama, C. S., & Wijesinghe, K. J. (2023). Bright NIR-emitting styryl pyridinium dyes with large stokes’ shift for sensing applications. Biosensors, 13(8), 799. https://doi.org/10.3390/bios13080799

Wu, L., Huang, J., Pu, K., & James, T. D. (2021). Dual-locked spectroscopic probes for sensing and therapy. Nature Reviews Chemistry, 5(6), 406-421. https://doi.org/10.1038/s41570-021-00277-2

Zhang, B., Chen, H., Shi, L., Guo, R., Wang, Y., Zheng, Y., ... & Zhang, X. (2024). Nitroreductase-based “turn-on” fluorescent probe for bacterial identification with visible features. Acs Sensors, 9(9), 4560-4567. https://doi.org/10.1021/acssensors.3c02785

Liu, J., Abdulkadir, A. Z., Wu, S., Gao, Y., Butuyuyu, B. J., Wong, K. M. C., ... & Zhang, P. (2025). Cyanine-scaffold fluorogenic probes for visual detection of nitroreductase in living bacteria. Journal of Materials Chemistry B, 13(38), 12224-12233. https://doi.org/10.1039/D5TB00758E

Morsby, J. J., Zhang, Z., Burchett, A., Datta, M., & Smith, B. D. (2024). Ratiometric near-infrared fluorescent probe for nitroreductase activity enables 3D imaging of hypoxic cells within intact tumor spheroids. Chemical Science, 15(10), 3633-3639. https://doi.org/10.1039/D3SC06058F

An, Z., Sun, Y., Wang, D., Ge, J., Zhao, C., Lu, H., ... & Wang, Q. (2024). Ultra-sensitive and fast nitroreductase-responsive fluorescence probe for real-time bioimaging of hypoxia in tumor cells and zebrafish wound inflammation. Sensors and Actuators B: Chemical, 420, 136458. https://doi.org/10.1016/j.snb.2024.136458

Sarkar, S., Shil, A., Jun, Y. W., Yang, Y. J., Cheng, R., Wu, W., ... & Ahn, K. H. (2025). Real-Time Detection of Reduced Nitroreductase with a Reversible Fluorescent Probe. Advanced Science, 12(42), e08689. https://doi.org/10.1002/advs.202508689

Gao, W., Shi, A., Hou, Y., Zhang, P., Zhang, Q., & Ding, C. (2025). A turn on fluorescent probe for nitroreductase activity and its application in real-time imaging of tumor hypoxia. Talanta, 290, 127804. https://doi.org/10.1016/j.talanta.2025.127804

Zhang, S., Ma, M., Zhao, C., Li, J., Xu, L., Zhang, Z., ... & Song, D. (2024). A novel low-background nitroreductase fluorescent probe for real-time fluorescence imaging and surgical guidance of thyroid cancer resection. Biosensors and Bioelectronics, 261, 116514. https://doi.org/10.1016/j.bios.2024.116514

Hu, C., Liu, H., Zhang, Z., Li, L., Mao, G. J., Cheng, W., & Zhou, L. (2025). A Self-Calibrating Fluorescent-Photoacoustic Integrated Probe Enables Fast Visualizing Pancreatic Cancer and Imaging-Guided Tumor Surgery. Small, 21(20), 2408527. https://doi.org/10.1002/smll.202408527