An Activatable Nitrobenzoselenadiazole-based Probe for Selective Protein Labeling and Targeted Phototherapy

doi:10.1007/s40242-026-6017-0

Abstract

Abstract: Developing fluorescent probes with high selectivity for target proteins in living cells remains a significant challenge. Conventional probes, such as nitrobenzoxadiazole (NBD) derivatives, often suffer from limited selectivity and short emission wavelengths, limiting their performance in complex biological environments. Here, we report the design and synthesis of NBSe-biotin, a red-emitting fluorescent probe based on the nitrobenzoselenadiazole (NBSe) scaffold. Compared to NBD analogues, NBSe-biotin exhibits red-shifted absorption and emission, enhanced chemical selectivity, and a turn-on fluorescence response upon specific binding to avidin, with emission at 604 nm. Live-cell imaging demonstrates that NBSe-biotin selectively visualizes overexpressed biotin receptors on HeLa cell membranes, with minimal background fluorescence in 293T control cells. Beyond imaging, this strategy enables the covalent attachment of small-molecule photosensitizers to proteins, and the receptor-mediated activation of NBSe-biotin phototoxicity in live cells illustrates its potential to incorporate diverse cancer biomarker-targeting ligands. Together, these features establish NBSe-biotin as a versatile platform for fluorescence-guided protein detection, live-cell imaging, and targeted phototherapy.

Key words: Nitrobenzoselenadiazole, Lysine, Avidin, Phototherapy

REN Xiaojie, YI Wenlong, YANG Li, YE Xinyu, LI Cheuk Hin, ZHANG Fan, LIN Jiafu, SUN Hongyan. An Activatable Nitrobenzoselenadiazole-based Probe for Selective Protein Labeling and Targeted Phototherapy[J]. Chemical Research in Chinese Universities, 2026, 42(2): 526-533.

References

[1] Yin X., Cheng Y., Feng Y., Stiles W. R., Park S. H., Kang H., Choi H. S., Adv. Drug. Deliv. Rev., 2022, 189, 114483.
[2] Li J., Kang M., Zhang Z., Li X., Xu W., Wang D., Gao X., Tang B. Z., Angew. Chem. Int. Ed., 2023, 62, e202301617.
[3] Wang Y., Ma K., Kang M., Yan D., Niu N., Yan S., Sun P., Zhang L., Sun L., Wang D., Tan H., Tang B. Z., Chem. Soc. Rev., 2024, 53, 12014.
[4] Zhou Z., Song J., Nie L., Chen X., Chem. Soc. Rev., 2016, 45, 6597.
[5] Sun B., Bte Rahmat J. N., Zhang Y., Biomaterials, 2022, 291, 121875.
[6] Zhang W., Li X., Kang M., Zhang Z., Pei Y., Fan M., Yan D., Zhang Y., Yang C., Xu G., Wang D., Xu Z., Tang B. Z., ACS Mater. Lett., 2024, 6, 2174.
[7] Wang M., Huang G., You Z., Jia R., Zhong Y., Bai F., Chem. Res. Chinese Universities, 2023, 39, 612.
[8] Ma F., Zhang R., Wang B., Liang Z., Zhang S., Jiang J., Tan H., Xing G., Kwok R. T. K., Lam J. W. Y., Zhao Z., Tang B. Z., J. Am. Chem. Soc., 2025, 147, 29815.
[9] An J., Tang S., Hong G., Chen W., Chen M., Song J., Li Z., Peng X., Song F., Zheng, W. H., Nat. Commun., 2022, 13, 2225.
[10] Li Y., Zhang D., Yu Y., Zhang L., Li L., Shi L., Feng G., Tang B. Z., ACS Nano, 2023, 17, 16993.
[11] Zhang P., Shen Q., Yang J., Zhao Z., Gao A., Chen S., Zhang Y., Meng L., Dang D., Matter, 2024, 7, 4342.
[12] Ma W., Wen P., Chen Z., Cetin M. M., Fang Z., Yang Z., Small, 2025, 21, 10047.
[13] Zhang J., Ma W., Luo H., Zhang K., Lv J., Jiang L., Huang Y., Song J., Yang Z., Huang W., Adv. Healthcare Mater., 2023, 13, 2303175.
[14] Wang Z., Zhou Y., Hao Y., Zhao, Z., Gao A., Ma H., Zhang P., Shen Q., Xu R., Xu Y., Dang D., Meng L., Nano Lett., 2024, 24, 3005.
[15] Chen P., Zhang G., Li J., Ma L., Zhou J., Zhu M., Li S., Wang Z., Chem. Res. Chinese Universities, 2024, 40, 293.
[16] Zhang Z., Xu W., Kang M., Wen H., Guo H., Zhang P., Xi L., Li K., Wang L., Wang D., Tang B. Z., Adv. Mater., 2020, 32, 2003210.
[17] Li J., Niu N., Wang D., Liu X., Qin Y., Wang L., Tang B. Z., Wang D., Sci. China Chem., 2024, 67, 2647.
[18] Gu H., Liu W., Sun W., Du J., Fan J., Peng X., Chem. Sci., 2022, 13, 9719.
[19] Shin J., Xu Y., Koo S., Lim J. H., Lee J. Y., Sharma A., Sun Y., Kim J. S., Matter, 2021, 4, 2508.
[20] Teng K. X., Niu L. Y., Xie N., Yang Q. Z., Nat. Commun., 2022, 13, 6179.
[21] Deng Y., Song P., Chen X., Huang Y., Hong L., Jin Q., Ji J., ACS Nano, 2020, 14, 9711.
[22] Liu Y., Zhang J., Zhou X., Wang Y., Lei S., Feng G., Wang D., Huang P., Lin J., Angew. Chem. Inter. Ed., 2024, 63, e202408064.
[23] Chen X., Shi L., Ran X. Y., Xu J. X., Zhang L. N., Kong Q. Q., Yu X. Q., Li K., Adv. Funct. Mater., 2024, 34, 2400728.
[24] Wang Y., Liao J., Lyu Y., Guo Q., Zhu Z., Wu X., Yu J., Wang Q., Zhu W. H., Adv. Funct. Mater., 2023, 33, 2301692.
[25] Wang Z., Ma W., Yang Z., Kiesewetter D. O., Wu Y., Lang L., Zhang G., Nakuchima S., Chen J., Su, Y., Han S., Wu L. G., Jin A. J., Huang W., J. Am. Chem. Soc., 2024, 146, 28973.
[26] Lin Y., Niu L. Y., Teng K. X., Yang Q. Z., J. Am. Chem. Soc., 2025, 147, 35172.
[27] Zhang D., Zhao Z., Tang Q., Jia H., Tang B. Z., Feng G., Angew. Chem. Inter. Ed., 2025, 64, e202516764.
[28] Hu X., Fang Z., Sun F., Zhu C., Jia M., Miao X., Huang L., Hu W., Fan Q., Yang Z., Huang W., Angew. Chem. Inter. Ed., 2024, 63, e202401036.
[29] Diao S., He X., Wu Y., Yin L., Huang Y., Zhou W., Xie C., Fan Q., Chem. Sci., 2025, 16, 10909.
[30] Zhang J., Luo H., Ma W., Lv J., Wang B., Sun F., Chi W., Fang Z., Yang Z., Adv. Sci., 2025, 12, 2500293.
[31] Yue Y., Li B., Wang D., Wu C., Li Z., Liu B., Adv. Funct. Mater., 2024, 35, 2414542.
[32] Li Y., Zhuang J., Lu Y., Li N., Gu M., Xia J., Zhao N., Tang B. Z., ACS Nano, 2021, 15, 20453.
[33] Wang H., Qin T., Wang W., Zhou X., Lin F., Liang G., Yang Z., Chi Z., Tang B. Z., Adv. Sci., 2023, 10, 2301902.
[34] Li C., Jiang G., Yu J., Ji W., Liu L., Zhang P., Du J., Zhan C., Wang J., Tang B. Z., Adv. Mater., 2022, 35, 2208229.
[35] Chen X., Mendes B. B., Zhuang Y., Conniot J., Mercado Argandona S., Melle F., Sousa D. P., Perl, D., Chivu A., Patra H. K., Shepard W., Conde J., Fairen-Jimenez D., J. Am. Chem. Soc., 2024, 146, 1644.
[36] Tian M., Zhao X., Zhao S., Li Y., Wang R., Guo J., Liu P., Chem. Commun., 2025, 61, 10985.
[37] Zheng P., Ding Y., Abdurahman A., Zhu J., Tian L., Liu Y., Liu Q., Wang L., Wang D., Tang B. Z., Sci. China Chem., 2025, 68, 3086.
[38] Li, J., Zhang Z., Jiang S., Kang M., Wang D., Li X., Huang P., Gao X., Wang D., Tang B. Z., Adv. Funct. Mater., 2024, 34, 2401627.
[39] Peng Y., Mo R., Yang M., Xie H., Ma F., Ding Z., Wu S., Lam J. W. Y., Du J., Zhang J., Zhao Z., Tang B. Z., ACS Nano, 2024, 18, 26140.
[40] Xie H., Liu W., Cheng S., Peng Y., Mo R., Zhang R., Cai J., Xin P., Tian Z., Kwok R. T. K., Lam J. W. Y., Xu F., Jin G., Zhang J., Tang B. Z., ACS Nano, 2025, 19, 37511.
[41] Yang Z., Zhang Z., Lei Z., Wang D., Ma H., Tang B. Z., ACS Nano, 2021, 15, 7328.