Ge Chun, Huang Xintong, Zhang Sujie, Yuan Man, Tan Zhaoyi, Xu Chen, Jie Qiong, Zhang Jingjing, Zou Jianjun, Zhu Yubing, Feng Dong, Zhang Yue, Aa Jiye
Department of Pharmacy, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, China.
Department of Clinical Pharmacology Lab, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210006, China.
Cancer Cell Int. 2023 Jan 31;23(1):14. doi: 10.1186/s12935-023-02853-6.
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.
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.
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.
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.
作为5-氟尿嘧啶（5-FU）的前体药物，口服的卡培他滨（CAP）在肝脏中初步转化为活性代谢物，然后在肠道中释放5-FU以发挥抗肿瘤活性。由于CAP的代谢变化在其活化过程中起关键作用，因此在体外永远不能使用单一类型的肠道或肝细胞来评估其药代动力学（PK）和药效学（PD）特性。因此，我们旨在建立一种新型体外系统，以有效评估这类前体药物的PK和PD。
通过在一个系统中同时使用结肠直肠癌（CRC）和肝癌细胞系建立共培养细胞模型。分别使用细胞计数试剂盒-8（CCK-8）和流式细胞术分析来评估细胞活力和凋亡。使用蛋白质免疫印迹分析测量凋亡相关蛋白表达水平。开发了一种选择性液相色谱-串联质谱（LC-MS/MS）方法用于共培养模型中的细胞PK。
CAP对五种单层CRC细胞系（SW480、LoVo、HCT-8、HCT-116和SW620）或肝癌细胞系（HepG2）几乎没有抗增殖作用。然而，在含有CRC和肝癌细胞系的共培养模型中，CAP对每种CRC细胞系都具有显著的抗肿瘤活性，尽管其对五种CRC细胞系的作用有所不同。此外，将CAP与HepG2细胞预孵育后，含有CAP活性代谢物的培养基对五种CRC细胞系也显示出抗肿瘤作用，表明肝细胞在CAP活化中起关键作用。
简单且经济高效的CRC和肝癌细胞共培养模型可以模拟前体药物在体内依赖于在肝脏中代谢转化为活性代谢物的过程，为在药物开发早期体外评估CAP类前体药物的PK和PD特性提供了一种有价值的策略。
