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有机阴离子转运多肽1B1(OATP1B1)是一种重要的肝摄取转运蛋白,介导许多内源性物质和药物转运进入肝细胞。为了鉴别和研究天然产物中潜在的OATP1B1调节剂,本实验采用稳定表达OATP1B1的中国仓鼠卵巢细胞(CHO-OATP1B1),在96孔板中测定了21种常见天然化合物和提取物对OATP1B1介导荧光素甲氨蝶呤转运的影响。该方法可进一步运用于对化合物库的高通量筛选。本研究表明,一些黄酮类化合物(如槲皮素、槲皮苷、芦丁、菊花梗叶总黄酮和桑皮黄素等)和三萜类化合物(如甘草次酸和甘草酸等)在体外对OATP1B1的功能有显著抑制作用,其IC50值均小于16μmol·L-1。甘草次酸的IC50值与其临床血药浓度相当,预示其有可能引起OATP1B1介导的药物-药物相互作用。构效关系分析表明,黄酮苷元比其相应的糖苷对OATP1B1的抑制作用更强,而且糖的类型和糖链长度对其活性均有影响。此外,以OATP1B1的底物氟伐他汀和罗伐他汀为探针药物,体外研究了几种天然化合物对OATP1B1功能影响的底物依赖性。结果表明,天然产物对OATP1B1转运功能的影响具有底物依赖性,有助于预测和避免潜在的OATP1B1介导的不良药物-药物和药物-食物相互作用,为临床合理用药提供实验依据和指导。
Organic anion transport polypeptide 1B1 (OATP1B1) is an important hepatic uptake transport protein that mediates the transport of many endogenous substances and drugs into hepatocytes. In order to identify and study potential OATP1B1 modulators in natural products, we tested the effects of 21 common natural compounds and extracts on OATP1B1-mediated fluorescence in 96-well plates using Chinese Hamster Ovary cells (OATP1B1) stably expressing OATP1B1 Effect of methotrexate transport. This method can be further applied to high-throughput screening of compound libraries. This study showed that some flavonoids (such as quercetin, quercitrin, rutin, chrysanthemum total flavonoids and mulberry flavin, etc.) and triterpenes (such as glycyrrhetinic acid and glycyrrhizin, etc.) in vitro OATP1B1 function was significantly inhibited, the IC50 values were less than 16μmol·L-1. IC50 value of glycyrrhetinic acid is comparable to its clinical plasma concentration, suggesting that it may cause OATP1B1-mediated drug-drug interaction. The structure-activity relationship analysis showed that flavonoid aglycone has a stronger inhibitory effect on OATP1B1 than its corresponding glycoside, and the type of sugar and the length of sugar chain have an impact on its activity. In addition, substrate dependence of OATP1B1 on the function of OATP1B1 was investigated in vitro using fluvastatin and lovastatin, OATP1B1 substrates. The results showed that the natural product had a substrate-dependent effect on OATP1B1 transport function, which could help predict and avoid potential OATP1B1-mediated adverse drug-drug-drug-food interactions and provide experimental evidence and guidance for clinical rational drug use.