School of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong Special Administrative Region.
Department of Microbiology, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong Special Administrative Region.
J Ethnopharmacol. 2019 Oct 28;243:112097. doi: 10.1016/j.jep.2019.112097. Epub 2019 Jul 17.
According to Traditional Chinese Medicine theory, influenza is categorized as a warm disease or Wen Bing. The Wen Bing formulas, such as Yin-Qiao-San and Sang-Ju-Yin, are still first-line herbal therapies in combating variant influenza virus. To continue our study on the pharmacokinetic and pharmacodynamic interactions between Wen Bing formulas and oseltamivir (OS), the first-line western drug for the treatment of influenza, further interactions between OS and the eight single herbs and their relevant marker components from Wen Bing formulas were investigated in the current study.
To establish an in-vitro screening platform for investigation of the potential anti-influenza herbs/herbal components that may have pharmacokinetic and pharmacodynamic interactions with OS.
To screen potential inhibition on OS hydrolysis, 1 μg/mL of OS is incubated with herbs/herbal components in diluted rat plasma, microsomes and human recombinant carboxylesterase 1(hCE1) under optimized conditions. MDCK-WT and MDCK-MDR1 cell lines are utilized to identify potential modification on P-gp mediated transport of OS by herbs/herbal components. Caco-2 cells with and without Gly-Sar inhibition are performed to study the uptake of OS via PEPT1 transporters. Modification on OAT3 mediated transport is verified by the uptake of OS on HEK293-MOCK/HEK293-OAT3 cells. Anti-virus effects were evaluated using plaque reduction assay on H1N1 and H3N2 viruses. Potential pharmacokinetic and pharmacodynamic interaction between OS (30 mg/kg) and the selected herb, Radix Scutellariae (RS), at 300-600 mg/kg were carried out on rats. All samples are analyzed by an LC/MS/MS method for the contents of OS and OSA. A mechanistic PK model was developed to interpret the HDI between OS and RS in rats.
Our developed platform was successfully applied to screen the eight herbal extracts and their ten marker components on metabolic inhibition of OS and modification of OS transport mediated by P-gp, OAT3 and PEPT1. Results from six in-vitro experiments were analyzed after converting raw data from each experiment to corresponding fold-change (FC) values, based on which Radix Scutellariae (RS) were selected to have the most HDI potential with OS. By analyzing the plasma and urine pharmacokinetic data after co-administration of OS with a standardized RS extract in rats using an integrated population pharmacokinetics model, it is suggested that RS could inhibit OS hydrolysis during absorption and increase the absorbed fraction of OS, which leads to the increased ratio of OS concentration versus that of OSA in both rat plasma and urine. Never the less, the anti-virus effects of 2.5 h post-dose rat plasma were not influenced by co-administration of OS with RS.
A six-dimension in-vitro screening platform has been developed and successfully applied to find RS as a potential herb that would influence the co-administrated OS in rats. Although co-administered RS could inhibit OS hydrolysis during absorption and increase the absorbed fraction of OS, which lead to the increased ratio of OS concentration versus that of OSA in both rat plasma and urine, the anti-virus effect of OS was not influenced by co-administered RS.
目的：建立一个体外筛选平台，用于研究与奥司他韦（OS）可能具有药代动力学和药效学相互作用的潜在抗流感草药/草药成分。
材料和方法：为了筛选对 OS 水解的潜在抑制作用，将 1μg/mL 的 OS 与草药/草药成分在稀释的大鼠血浆、微粒体和人重组羧酸酯酶 1（hCE1）中在优化条件下孵育。利用 MDCK-WT 和 MDCK-MDR1 细胞系鉴定草药/草药成分对 P-糖蛋白介导的 OS 转运的潜在修饰作用。通过 Caco-2 细胞上的 Gly-Sar 抑制作用研究 OS 通过 PEPT1 转运体的摄取。通过在 HEK293-MOCK/HEK293-OAT3 细胞上摄取 OS 来验证 OAT3 介导的转运的修饰作用。采用 H1N1 和 H3N2 病毒的蚀斑减少测定法评价抗病毒效果。在大鼠中进行 OS（30mg/kg）与选定草药黄芩（RS）的 300-600mg/kg 的潜在药代动力学和药效学相互作用研究。所有样品均采用 LC/MS/MS 法分析 OS 和 OSA 的含量。建立了一个机制性 PK 模型，以解释 OS 和 RS 在大鼠中的 HDI。
结果：我们开发的平台成功地应用于筛选八种草药提取物及其十种标记成分对 OS 代谢抑制作用以及 P-糖蛋白、OAT3 和 PEPT1 介导的 OS 转运的修饰作用。在将每个实验的原始数据转换为相应的倍数变化（FC）值后，对来自六个体外实验的数据进行了分析，基于此，黄芩（RS）被选为与 OS 具有最大 HDI 潜力的草药。通过在大鼠中使用集成群体药代动力学模型分析 OS 与标准化 RS 提取物共同给药后血浆和尿液中的药代动力学数据，提示 RS 可能在吸收过程中抑制 OS 水解，并增加 OS 的吸收分数，从而导致大鼠血浆和尿液中 OS 浓度与 OSA 浓度的比值增加。尽管如此，在给药后 2.5 小时，大鼠血浆的抗病毒作用不受 OS 与 RS 共同给药的影响。
结论：建立了一个六维体外筛选平台，并成功应用于发现黄芩（RS）是一种可能影响与 OS 共同给药的潜在草药。虽然共同给予 RS 可能会抑制 OS 在吸收过程中的水解并增加 OS 的吸收分数，从而导致大鼠血浆和尿液中 OS 浓度与 OSA 浓度的比值增加，但 OS 的抗病毒作用不受共同给予 RS 的影响。
