Marsdenia tenacissima extract inhibits gefitinib metabolism in vitro by interfering with human hepatic CYP3A4 and CYP2D6 enzymes
Marsdenia tenacissima extract inhibits gefitinib metabolism in vitro by interfering with human hepatic CYP3A4 and CYP2D6 enzymes
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Marsdenia tenacissima 提取物通过干扰人肝 CYP3A4 和 CYP2D6 酶来抑制吉非替尼体外代谢。
DOI：
10.1016/j.jep.2013.10.021
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发表时间：
2014-01-10
影响因子：
5.4
通讯作者：
Li, Ping-Ping
中科院分区：
文献类型：
--
作者：
Han, Shu-Yan;Zhao, Hai-Yu;Li, Ping-Ping
民族药理学相关性：Marsdenia tenacissima (Roxb.) Wight et Am 的茎，主要产于中国云南，在中国长期以来被用作治疗癌症的药物。消癌平注射液是马斯登尼亚茎的水溶性部分，几十年来作为抗癌剂在临床使用。我们之前的研究表明，Marsdenia tenacissima 提取物 (MTE) 可恢复吉非替尼耐药的非小细胞肺癌 (NSCLC) 细胞对吉非替尼的敏感性，但其机制尚不清楚。吉非替尼主要通过人细胞色素 P450 (CYP) 3A4 和 CYP2D6 酶进行肝脏代谢。本研究旨在评估 MTE 是否通过人肝 P450 酶干扰吉非替尼代谢。 材料和方法：通过将混合的人肝微粒体与 CYP1A2、CYP2C9、CYP2C19、CYP2D6 和 CYP3A4 的特定底物探针在不存在和存在的情况下孵育，采用鸡尾酒底物测定法来测试 MTE 对主要 CYP 酶活性的影响。 MTE。使用重组人 CYP450 酶来预测在不存在和存在 MTE 的情况下吉非替尼的体外代谢清除率。通过高效液相色谱串联质谱（HPLC-MS/MS）检测底物探针和吉非替尼的代谢物。采用人肝癌HepG2细胞，考察吉非替尼单用或联合MTE对CYP3A4和CYP2D6 mRNA及蛋白表达的影响。结果：鸡尾酒底物实验显示，MTE抑制人肝微粒体CYP450活性，抑制率依次为3A4>2C9>2C19>1A2>2D6。 MITE 与吉非替尼联合给药显着降低吉非替尼对 CYP2D6 和 CYP3A4 的体外内在清除率 (Cl-int) 分别降低 2.6 倍和 4.0 倍，但不影响其他 CYP450。在吉非替尼和MTE联合治疗中，人肝癌HepG2细胞中CYP2D6和CYP3A4 mRNA和蛋白表达大大降低。结论：我们证明MTE通过干扰CYP3A4和CYP2D6来抑制吉非替尼代谢。同时，MTE联合吉非替尼下调HepG2细胞中CYP3A4和CYP2D6的mRNA和蛋白表达。因此，这些数据表明 MTE 是一种有前途的草药，可以通过改善其代谢稳定性来增强吉非替尼的疗效。 (C) 2013 年，爱思唯尔爱尔兰有限公司出版。
Ethnopharmacological relevance: The stem of Marsdenia tenacissima (Roxb.) Wight et Am, is mainly produced in Yunnan China and has long been used as a medicine to treat cancer in China. Xiao-Ai-Ping injection, the water-soluble part of the stem of Marsdenia tenacissima, is administrated as an anti-cancer agent in clinics for decades. Our previous study showed that Marsdenia tenacissima extract (MTE) restored gefitinib sensitivity in gefitinib-resistant non-small cell lung cancer (NSCLC) cells, but the mechanism involved is unknown. Gefitinib undergoes hepatic metabolism predominantly through human cytochrome P450 (CYP) 3A4 and CYP2D6 enzymes. This study aims to evaluate whether MTE interferes with gefitinib metabolism via human hepatic P450 enzymes.Material and methods: A cocktail-substrate assay was used to test the effect of MTE on major CYP enzyme activities by incubation of pooled human liver microsomes with specific substrate probes of CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 in the absence and presence of MTE. Recombinant human CYP450 enzymes were used to predict in vitro gefitinib metabolic clearance in the absence and presence of MTE. The metabolites of the substrate probes and gefitinib were detected by high-performance liquid chromatographic tandem mass spectrometry (HPLC-MS/MS). Human hepatoma HepG2 cells were used to investigate the effect of gefitinib alone or in combination with MTE on CYP3A4 and CYP2D6 mRNA and protein expression.Results: The cocktail-substrate assay showed that MTE inhibited CYP450 activities in human liver microsomes with the inhibition rate of 3A4> 2C9 > 2C19> 1A2 > 2D6. The co-administration of MITE with gefitinib significantly decreased the in vitro intrinsic clearance (Cl-int) of gefitinib by 2.6 and 4.0-fold for CYP2D6 and CYP3A4, respectively, but did not affect other CYP450s. CYP2D6 and CYP3A4 mRNA and protein expression in human hepatoma HepG2 cells were greatly reduced in the combined gefitinib and MTE treatment.Conclusion: We demonstrate that MTE inhibits gefitinib metabolism by interfering with CYP3A4 and CYP2D6. Meanwhile, MTE combined with gefitinib down-regulates the mRNA and protein expression of CYP3A4 and CYP2D6 in the HepG2 cells. Thus, these data suggest that MTE is a promising herbal medicine to enhance gefitinib efficacy through improving its metabolic stability. (C) 2013 Published by Elsevier Ireland Ltd.