Reactive Cysteines in Proteins are the Dominant Reductants for Platinum(IV) Prodrug Activation in Live Cells

Shu Chen; Wang Peng; Houzong Yao; Zhiqin Deng; Zhao Yue; Gongyuan Liu; Jiaqian Xu; Naixin Lin; Weikang Xu; Jianbo Yue; Guangyu Zhu

doi:10.1002/anie.202416396

Reactive Cysteines in Proteins are the Dominant Reductants for Platinum(IV) Prodrug Activation in Live Cells

Shu Chen, Wang Peng, Houzong Yao, Zhiqin Deng, Zhao Yue, Gongyuan Liu, Jiaqian Xu, Naixin Lin, Weikang Xu, Jianbo Yue, Guangyu Zhu^*

^*Corresponding author for this work

Research output: Journal Publications and Reviews › RGC 21 - Publication in refereed journal › peer-review

4 Citations (Scopus)

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Abstract

The intracellular reduction of Pt(IV) prodrugs is crucial for their anticancer efficacy. However, the major components responsible for the reduction of Pt(IV) complexes within live cells remain elusive. Herein, we developed an aminoluciferin-functionalized Pt(IV) complex, Pt-Luc, that can be used as a bioluminescent reporter for real-time monitoring of Pt(IV) reduction in live cancer cells by capturing immediate bioluminescent signals from the released aminoluciferin. Utilizing this powerful reporter, we found that the reduction of Pt(IV) prodrugs in live cancer cells significantly slows down when cysteine levels are reduced, while the levels of glutathione do not impact the reduction rate. Further investigation reveals that reactive cysteines in proteins, rather than small-molecule thiols, play a primary role in reducing the Pt(IV) complex. In vivo studies reveal a substantial 63% decrease in bioluminescence from Pt-Luc in thiol-blocking tumors in mice, reinforcing the pivotal role of reactive cysteines in Pt(IV) reduction. This study provides valuable insights into the activation mechanisms of Pt(IV) prodrugs in live cells and in vivo, enhancing our understanding of prodrug activation beyond buffer systems or fixed cells. © 2025 The Author(s). Angewandte Chemie International Edition published by Wiley-VCH GmbH.

Original language	English
Article number	e202416396
Journal	Angewandte Chemie International Edition
Volume	64
Issue number	25
Online published	11 Apr 2025
DOIs	https://doi.org/10.1002/anie.202416396
Publication status	Published - 17 Jun 2025

Funding

The authors would like to thank the Hong Kong Research Grants Council (Grant Nos. CityU 11303320, CityU 11302221, CityU 11313222, CityU 11304923, 11306724, and C1018\u201023G), the National Natural Science Foundation of China (Grant Nos. 22077108 and 22277103), the Science Technology and Innovation Committee of Shenzhen Municipality (JCYJ20210324120004011), and the City University of Hong Kong (Grant No. 7020014) for funding support.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Research Keywords

Bioluminescence imaging
Live cells
Platinum(IV) prodrug
Platinum(IV) reduction
Prodrug activation

Publisher's Copyright Statement

This full text is made available under CC-BY 4.0. https://creativecommons.org/licenses/by/4.0/

RGC Funding Information

RGC-funded

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10.1002/anie.202416396

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title = "Reactive Cysteines in Proteins are the Dominant Reductants for Platinum(IV) Prodrug Activation in Live Cells",

abstract = "The intracellular reduction of Pt(IV) prodrugs is crucial for their anticancer efficacy. However, the major components responsible for the reduction of Pt(IV) complexes within live cells remain elusive. Herein, we developed an aminoluciferin-functionalized Pt(IV) complex, Pt-Luc, that can be used as a bioluminescent reporter for real-time monitoring of Pt(IV) reduction in live cancer cells by capturing immediate bioluminescent signals from the released aminoluciferin. Utilizing this powerful reporter, we found that the reduction of Pt(IV) prodrugs in live cancer cells significantly slows down when cysteine levels are reduced, while the levels of glutathione do not impact the reduction rate. Further investigation reveals that reactive cysteines in proteins, rather than small-molecule thiols, play a primary role in reducing the Pt(IV) complex. In vivo studies reveal a substantial 63% decrease in bioluminescence from Pt-Luc in thiol-blocking tumors in mice, reinforcing the pivotal role of reactive cysteines in Pt(IV) reduction. This study provides valuable insights into the activation mechanisms of Pt(IV) prodrugs in live cells and in vivo, enhancing our understanding of prodrug activation beyond buffer systems or fixed cells. {\textcopyright} 2025 The Author(s). Angewandte Chemie International Edition published by Wiley-VCH GmbH.",

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T1 - Reactive Cysteines in Proteins are the Dominant Reductants for Platinum(IV) Prodrug Activation in Live Cells

AU - Chen, Shu

AU - Peng, Wang

AU - Yao, Houzong

AU - Deng, Zhiqin

AU - Yue, Zhao

AU - Liu, Gongyuan

AU - Xu, Jiaqian

AU - Lin, Naixin

AU - Xu, Weikang

AU - Yue, Jianbo

AU - Zhu, Guangyu

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N2 - The intracellular reduction of Pt(IV) prodrugs is crucial for their anticancer efficacy. However, the major components responsible for the reduction of Pt(IV) complexes within live cells remain elusive. Herein, we developed an aminoluciferin-functionalized Pt(IV) complex, Pt-Luc, that can be used as a bioluminescent reporter for real-time monitoring of Pt(IV) reduction in live cancer cells by capturing immediate bioluminescent signals from the released aminoluciferin. Utilizing this powerful reporter, we found that the reduction of Pt(IV) prodrugs in live cancer cells significantly slows down when cysteine levels are reduced, while the levels of glutathione do not impact the reduction rate. Further investigation reveals that reactive cysteines in proteins, rather than small-molecule thiols, play a primary role in reducing the Pt(IV) complex. In vivo studies reveal a substantial 63% decrease in bioluminescence from Pt-Luc in thiol-blocking tumors in mice, reinforcing the pivotal role of reactive cysteines in Pt(IV) reduction. This study provides valuable insights into the activation mechanisms of Pt(IV) prodrugs in live cells and in vivo, enhancing our understanding of prodrug activation beyond buffer systems or fixed cells. © 2025 The Author(s). Angewandte Chemie International Edition published by Wiley-VCH GmbH.

AB - The intracellular reduction of Pt(IV) prodrugs is crucial for their anticancer efficacy. However, the major components responsible for the reduction of Pt(IV) complexes within live cells remain elusive. Herein, we developed an aminoluciferin-functionalized Pt(IV) complex, Pt-Luc, that can be used as a bioluminescent reporter for real-time monitoring of Pt(IV) reduction in live cancer cells by capturing immediate bioluminescent signals from the released aminoluciferin. Utilizing this powerful reporter, we found that the reduction of Pt(IV) prodrugs in live cancer cells significantly slows down when cysteine levels are reduced, while the levels of glutathione do not impact the reduction rate. Further investigation reveals that reactive cysteines in proteins, rather than small-molecule thiols, play a primary role in reducing the Pt(IV) complex. In vivo studies reveal a substantial 63% decrease in bioluminescence from Pt-Luc in thiol-blocking tumors in mice, reinforcing the pivotal role of reactive cysteines in Pt(IV) reduction. This study provides valuable insights into the activation mechanisms of Pt(IV) prodrugs in live cells and in vivo, enhancing our understanding of prodrug activation beyond buffer systems or fixed cells. © 2025 The Author(s). Angewandte Chemie International Edition published by Wiley-VCH GmbH.

KW - Bioluminescence imaging

KW - Live cells

KW - Platinum(IV) prodrug

KW - Platinum(IV) reduction

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ER -

Reactive Cysteines in Proteins are the Dominant Reductants for Platinum(IV) Prodrug Activation in Live Cells

Abstract

Funding

UN SDGs

Research Keywords

Publisher's Copyright Statement

RGC Funding Information

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GRF: Synthesis and Biological Evaluation of Near-Infrared-II Activatable Platinum Anticancer Prodrugs

CRF: Controllable Activation of Anticancer Prodrugs in vivo

GRF: Plasma Membrane-anchored Platinum(IV) Anticancer Photooxidants to Reduce Drug Resistance

Cite this

Reactive Cysteines in Proteins are the Dominant Reductants for Platinum(IV) Prodrug Activation in Live Cells

Abstract

Funding

UN SDGs

Research Keywords

Publisher's Copyright Statement

RGC Funding Information

Access Output

Other Access Options

Permanent Link

Other Files and Links

Fingerprint

Projects

GRF: Synthesis and Biological Evaluation of Near-Infrared-II Activatable Platinum Anticancer Prodrugs

CRF: Controllable Activation of Anticancer Prodrugs in vivo

GRF: Plasma Membrane-anchored Platinum(IV) Anticancer Photooxidants to Reduce Drug Resistance

Cite this