Citation: Shiwen Song, Mingyang Zhao, Xiangrong Song, Huaidong Yu, Xin Xu, Dong Sun, Jingkai Gu. Unraveling metabolic complexity of multi-payload PEG-irinotecan prodrug: A deconvolution-based LC-Q-TOF MS approach for preclinical pharmacokinetic characterization[J]. Chinese Chemical Letters, ;2026, 37(2): 112099. doi: 10.1016/j.cclet.2025.112099 shu

Unraveling metabolic complexity of multi-payload PEG-irinotecan prodrug: A deconvolution-based LC-Q-TOF MS approach for preclinical pharmacokinetic characterization

Shiwen Song ^{c, 1} ,
Mingyang Zhao ^{a, 1} ,
Xiangrong Song ^e ,
Huaidong Yu ^f ,
Xin Xu ^a ,
Dong Sun ^{a, *,} ,
Jingkai Gu ^{a, b, d, *,}

a.
Research Center for Drug Metabolism, School of Life Science, Jilin University, Changchun 130012, China
b.
State Key Laboratory of Supramolecular Structure and Materials, Center for Supramolecular Chemical Biology, College of Chemistry, Jilin University, Changchun 130012, China
c.
School of Pharmaceutical Science, Hainan University, Haikou 570100, China
d.
Beijing Institute of Drug Metabolism, Beijing 102209, China
e.
State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China
f.
Beijing GeZhu Biotechnology Co., Ltd., Beijing 102209, China

sun_dong@jlu.edu.cn

gujk@jlu.edu.cn

Received Date: 11 July 2025
Revised Date: 10 November 2025
Accepted Date: 11 November 2025
Available Online: 12 November 2025

Figures(5)

PEGylation, the controlled covalent conjugation of polyethylene glycol to therapeutics, enhances therapeutic efficacy through optimized pharmacokinetics. However, to date no high-molecular-weight PEGylated small-molecule prodrugs have received regulatory approval. This technological gap can be partially attributed to the exponential proliferation of metabolic intermediates resulting from multi-payload conjugation strategies, which introduces unprecedented analytical complexities in metabolite profiling and pharmacokinetic characterization. To address this challenge, we developed a liquid chromatography-triple-quadrupole/time-of-flight mass spectrometry platform for PEG20k-(irinotecan)₃, a Phase Ⅲ clinical candidate. Our methodology employs payload stoichiometry-based chromatographic resolution for clustering isomeric PEG species. Complementarily, diagnostic product ions at m/z 699.83, 569.27, and 587.28 enable systematic differentiation between double-loaded, single-loaded, and released irinotecan payload. This approach successfully identifies eight metabolic clusters spanning from PEG-conjugates, cleaved PEG segments, and released small-molecule species. Its demonstrated capacity to deconvolute complex metabolic profiles—through payload-stoichiometry based chromatographic resolution coupled with diagnostic ion analysis—positions this workflow as an attractive tool for accelerating the development of PEGylated small-molecule therapeutics.
- Pharmacokinetics,
- Multi-payload PEG-prodrug,
- High-molecular-weight PEG,
- LC-Q-TOF MS,
- Payload-based resolution
1. [1]
  Y. Gao, M. Joshi, Z. Zhao, S. Mitragotri, Bioeng. Transl. Med. 9 (2024) e10600. doi: 10.1002/btm2.10600
2. [2]
  F.M. Veronese, G. Pasut, Drug Discov. Today 10 (2005) 1451–1458. doi: 10.1016/S1359-6446(05)03575-0
3. [3]
  S. Yang, B. Chen, J. Wang, K. Zhang, Anal. Chem. 92 (2020) 8584–8590. doi: 10.1021/acs.analchem.0c01567
4. [4]
  M. Swierczewska, K.C. Lee, S. Lee, Expert Opin. Emerg. Drugs 20 (2015) 531–536. doi: 10.1517/14728214.2015.1113254
5. [5]
  H. Li, M. Zhang, J. He, et al., Chin. Chem. Lett. 36 (2025) 110615. doi: 10.1016/j.cclet.2024.110615
6. [6]
  W. Li, P. Zhan, E. De Clercq, H. Lou, X. Liu, Prog. Polym. Sci. 38 (2013) 421–444. doi: 10.1016/j.progpolymsci.2012.07.006
7. [7]
  E.J. Park, J. Choi, K.C. Lee, D.H. Na, Expert Opin. Emerg. Drugs 24 (2019) 107–119. doi: 10.1080/14728214.2019.1604684
8. [8]
  H. Park, A. Otte, K. Park, J. Control. Release 342 (2022) 53–65. doi: 10.1016/j.jconrel.2021.12.030
9. [9]
  M. Daniali, S. Nikfar, M. Abdollahi, Expert Opin. Pharmacother. 21 (2020) 883–891. doi: 10.1080/14656566.2020.1735354
10. [10]
  FDANonclinical Studies for the Safety Evaluation of Pharmaceutical Excipients (Guidance), 2005 https://www.fda.gov/regulatory-information/search-fda-guidance-documents/nonclinical-studies-safety-evaluationpharmaceutical-excipients.
11. [11]
  EMA-CHMPGuideline on the Pharmacokinetic and Clinical Evaluation of Modified Release Dosage Forms (EMA/CPMP/EWP/280/96 Corr1), 2015 https://www.ema.europa.eu/en/documents/presentation/presentation-emaegaworkshop-guideline-pharmacokinetic-and-clinical-evaluation-modifiedrelease-dosage-forms-beata-stepniewska_en.pdf.
12. [12]
  S. Song, D. Sun, H. Wang, et al., Acta Pharm. Sin. B 13 (2023) 3444–3453. doi: 10.1016/j.apsb.2023.01.011
13. [13]
  Z. Zhang, Y. Zhang, S. Song, et al., J. Sep. Sci. 43 (2020) 1978–1997. doi: 10.1002/jssc.201901340
14. [14]
  X. Zhou, X. Meng, L. Cheng, et al., Anal. Chem. 89 (2017) 5193–5200. doi: 10.1021/acs.analchem.6b04058
15. [15]
  X. Meng, Z. Zhang, J. Tong, et al., Acta Pharm. Sin. B 11 (2021) 1003–1009. doi: 10.1016/j.apsb.2021.02.018
16. [16]
  T. Wang, Y. Guo, Y. He, et al., Acta Pharm. Sin. B 10 (2020) 2002–2009. doi: 10.1016/j.apsb.2020.02.001
17. [17]
  L. Yin, C. Su, T. Ren, et al., Analyst 142 (2017) 4279–4288. doi: 10.1039/C7AN00470B
18. [18]
  T. Ren, R. Li, L. Zhao, et al., Acta Pharm. Sin. B 12 (2022) 3156–3166. doi: 10.1016/j.apsb.2022.01.014
19. [19]
  Z. Zhang, H. Jiang, Y. Zhang, et al., Anal. Chem. 92 (2020) 5978–5985. doi: 10.1021/acs.analchem.0c00189
20. [20]
  R. Bai, D. Sun, Y. Shan, et al., Chin. Chem. Lett. 32 (2021) 3237–3240. doi: 10.1016/j.cclet.2021.04.031
21. [21]
  L. Yin, T. Ren, S. Zhao, M. Shi, J. Gu, Talanta 206 (2020) 120184. doi: 10.1016/j.talanta.2019.120184
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2. [2]
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16. [16]
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