基于药效物质组及肝肾转运体排泄网络研究生脉方“君臣佐”配伍理论的药动学机制

Prescription compatibility is the core for Chinese medicinal formulae and exploring the variation rules of the active ingredient groups after combination based on pharmacokinetic perspective by using modern methods will be very crucial. A series of transporters distribute in hepatic and renal excretory system and can transport the complex constituents of Traditional Chinese Medicine (TCM). These constituents from TCM maybe also influence the activities of above transporters. The compatibility theory of ShengMai Formula (SMF) may imply a synergistic combination mechanism that competitions or regulations may occur among the active ingredient groups from different herbs in SMF mediated by hepatic and renal transporters network and result in the decrease of drug excretion and the enhancement of drug action. Our previous studies indicate that Rb1 and Rd significantly inhibit the uptake of ophiopogonin D (OPD) in OATP1B1/1B3-HEK293T cells, and OPD’s plasma concentrations in rats obviously rise after combination with Rb1 and Rd, which suggests there maybe some interactions among the complex constituents in ShenMai Injection mediated by OATPs. In this project, therefore, we will construct a series of overexpressing hepatic or renal transporter cell models and systematacially study the transporting mechanisms of the principal constitutions of Shengmai Formula mediated by hepatic and renal transporters network. Their impacts on transporters’ functions and the variations of their transporting process after compatibility also need to be explored. Furthermore, we also will investigate the profiles of distribution of target ingredients in blood and liver and its elimination from bile and urine ducts in rats when single or combined administration. The purpose is to illustrate the compatibility mechanisms of different active ingredient groups from “Monarch, Minister and Assistant” in SMF and its scientific connotation in terms of hepatic and renal transporter network.

方剂核心是药物配伍，采用现代方法从药动学角度探究中药配伍后药效物质组的变化规律是关键。在肝肾排泄系统中分布着复杂的药物转运体网络，中药复杂组分可以经肝肾转运排泄或对转运功能产生影响。生脉方配伍理论中可能蕴含着不同药味间的药效物质组发生基于肝肾转运体网络介导的竞争，导致排泄减少、药效增强的协同配伍机制。前期证实参麦方中Rb1和Rd显著抑制OATP1B1/1B3-HEK293T细胞中OPD的摄取，联用Rb1+Rd后大鼠体内OPD血药浓度明显升高，提示参麦方复杂组分间存在基于OATPs介导的相互作用。课题拟构建系列高表达肝肾转运体的细胞模型，系统研究生脉方主要药效物质组在肝肾转运体网络介导下的转运情况、对转运功能的影响及其配伍前后转运变化；研究单独与配伍给药后大鼠体内目标成分群的血肝分布、胆尿排泄及其对转运体表达调控，从肝肾转运体排泄网络角度阐明生脉方“君臣佐”配伍理论的科学内涵及药动学机制。

我们系统研究了生脉方主要活性组分对OATP1B1、OATP1B3、NTCP、BCRP、OAT1、OCT2转运功能的影响及其相互作用，发现甲基麦冬二氢高异黄酮A、甲基麦冬二氢高异黄酮B、麦冬皂苷D和麦冬皂苷D′对OATP1B1和OATP1B3的转运活性有影响；人参皂苷Rb1和Rd可显著抑制由OATPs/Oatps介导的麦冬皂苷D摄取；五味子甲素和五味子乙素可能是OATP1B1潜在的底物；人参皂苷Rg1、麦冬皂苷D′和五味子甲素是NTCP的潜在抑制剂，人参皂苷Re和五味子甲素B是 NTCP的潜在底物；人参总皂苷、五味子总木脂素、人参皂苷Rd、Rb1和五味子醇乙是BCRP的潜在抑制剂，人参皂苷Re和Rg1、甲基麦冬皂苷B和五味子甲素是BCRP的潜在底物；人参皂苷Re、人参皂苷Rd和甲基麦冬二氢高异黄酮A是OAT1的抑制剂，人参皂苷Re、人参皂苷 Rd 和甲基麦冬二氢高异黄酮A 可抑制五味子乙素经OAT1的摄取转运；人参皂苷Rb1和MA可能是OCT2的潜在底物。生脉方主要活性组分间存在基于上述转运体介导的相互作用及配伍机制。本课题从肝肾转运体排泄网络角度揭示了生脉方“君臣佐”配伍理论的科学内涵及药动学机制，为生脉方中药现代化研究及临床安全用药提供实验依据。