In vitro co-culture systems of hepatic and intestinal cells for cellular pharmacokinetic and pharmacodynamic studies of capecitabine against colorectal cancer

  • Cancer Cell Int. 2023 Jan 31;23(1):14. doi: 10.1186/s12935-023-02853-6.
Chun Ge  #  1  2  3 ,  Xintong Huang  #  3 ,  Sujie Zhang  #  3 ,  Man Yuan  3 ,  Zhaoyi Tan  4 ,  Chen Xu  4 ,  Qiong Jie  1  2  3 ,  Jingjing Zhang  1  2  3 ,  Jianjun Zou  1  2  3 ,  Yubing Zhu  5  6  7 ,  Dong Feng  8 ,  Yue Zhang  9 ,  Jiye Aa  4
Affiliations
  • 1. Department of Pharmacy, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, China.
  • 2. Department of Clinical Pharmacology Lab, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, China.
  • 3. Department of Clinical Pharmacy, School of Basic Medicine & Clinical Pharmacy, China Pharmaceutical University, Nanjing, 211198, China.
  • 4. Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, 210009, China.
  • 5. Department of Pharmacy, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, China. [email protected].
  • 6. Department of Clinical Pharmacology Lab, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, China. [email protected].
  • 7. Department of Clinical Pharmacy, School of Basic Medicine & Clinical Pharmacy, China Pharmaceutical University, Nanjing, 211198, China. [email protected].
  • 8. Nanjing Southern Pharmaceutical Technology Co., Ltd., Nanjing, 211100, China. [email protected].
  • 9. Key Laboratory of Drug Metabolism and Pharmacokinetics, State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing, 210009, China. [email protected].
  • # Contributed equally.
PMID: 36717845 DOI: 10.1186/s12935-023-02853-6
Abstract

Background: As a prodrug of 5-fluorouracil (5-FU), orally administrated capecitabine (CAP) undergoes preliminary conversion into active metabolites in the liver and then releases 5-FU in the gut to exert the anti-tumor activity. Since metabolic changes of CAP play a key role in its activation, a single kind of intestinal or hepatic cell can never be used in vitro to evaluate the pharmacokinetics (PK) and pharmacodynamics (PD) nature. Hence, we aimed to establish a novel in vitro system to effectively assess the PK and PD of these kinds of prodrugs.

Methods: Co-culture cellular models were established by simultaneously using colorectal Cancer (CRC) and hepatocarcinoma cell lines in one system. Cell Counting Kit-8 (CCK-8) and flow cytometric analysis were used to evaluate cell viability and Apoptosis, respectively. Apoptosis-related protein expression levels were measured using western blot analysis. A selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for cellular PK in co-culture models.

Results: CAP had little anti-proliferative effect on the five monolayer CRC cell lines (SW480, LoVo, HCT-8, HCT-116 and SW620) or the hepatocarcinoma cell line (HepG2). However, CAP exerted marked anti-tumor activities on each of the CRC cell lines in the co-culture models containing both CRC and hepatocarcinoma cell lines, although its effect on the five CRC cell lines varied. Moreover, after pre-incubation of CAP with HepG2 cells, the culture media containing the active metabolites of CAP also showed an anti-tumor effect on the five CRC cell lines, indicating the crucial role of hepatic cells in the activation of CAP.

Conclusion: The simple and cost‑effective co-culture models with both CRC and hepatocarcinoma cells could mimic the in vivo process of a prodrug dependent on metabolic conversion to active metabolites in the liver, providing a valuable strategy for evaluating the PK and PD characteristics of CAP-like prodrugs in vitro at the early stage of drug development.

Keywords
Capecitabine; Cellular pharmacodynamics; Cellular pharmacokinetics; Co-culture; Prodrug.
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