热淋清颗粒与喹诺酮类抗生素联用产生药物相互作用的药代动力学机制

Relinqing granule is a sole preparation made from Sijihong (Polygonum capitatum) which is one of the six ethnic Miao’s commonly used herb in Guizhou Province. Relinqing is often co-administered with quinolones against urinary tract infections with a unique therapeutic effect. The previous study (NSFC No. 81260688) indicated that CYP2C9 and CYP3A4 were induced by Sijihong and the main pharmacokinetic parameters of ciprofloxacin in vivo were significantly altered by Relinqing, which demonstrated the occurrence of drug-drug interaction (DDI) between them. Based on the point that the ADME process is the fundament of efficacy and DDI in vivo, the study on pharmacokinetics DDI will be carried out in the process of absorption, distribution, metabolism, excretion in order to reveal the nature and intensity of DDI and the action of the effect under the combination. The study will also put focus on the regulation of metabolic enzymes and transporters to explore the molecular pharmacokinetic mechanism mediated by CYP450s and transporters in the DDI. The project of the study will contribute to clarifying the association with the pharmacokinetics DDI and will be helpful for rational and scientific provision the theoretical basis of the combination of Relinqing and quinolones in clinical practice.

热淋清颗粒为贵州“六大苗药”之一四季红的单方制剂，临床常与喹诺酮类药物联合用于治疗尿路感染疗效显著。申请者在国家自然基金“贵州苗药四季红的吸收机制及体内过程研究”（81260688）中发现，四季红能诱导CYP2C9 与CYP3A4酶活性，合用热淋清颗粒可使环丙沙星在大鼠体内的药动学参数发生明显变化，提示两者发生了药动学药物相互作用（DDI）。本课题基于体内ADME过程为药物发挥疗效和DDI的基础，拟开展两药合用在体内吸收、分布、代谢、排泄过程中的药动学DDI，揭示两药发生DDI的性质与强度及作用环节，并重点研究代谢酶和转运体在DDI过程中的调控作用，探讨基于CYP450酶和转运体介导的DDI分子药代动力学机制，明确两药合用的机制与其药动学DDI的关联性，为阐明热淋清颗粒与喹诺酮类药物联用应用的临床合理性和科学内涵提供理论依据。

热淋清颗粒是以贵州“六大苗药”之一的头花蓼为原料制成的单方制剂，为国家中药保护品种和2020版《中国药典》收载品种。热淋清颗粒和喹诺酮类药物的联合应用已成为临床治疗尿路感染的常见治疗方式，但两药联用是否发生药物相互作用尚不清楚。本项目基于大鼠肾盂肾炎疾病模型，研究了热淋清颗粒在正常和肾盂肾炎大鼠体内的药动学及组织分布过程。基于整体药动学，研究了热淋清颗粒对左氧氟沙星在药动学、吸收、组织分布、排泄状况以及血浆蛋白结合率的影响。研究发现两药联用后发生了明显的相互作用，热淋清颗粒能显著减少左氧氟沙星在体内的暴露量，主要体现在大鼠血浆、组织器官和主要的排泄途径中。采用Caco-2细胞、大鼠外翻肠囊、在体循环肠灌流模型研究比较了左氧氟沙星与热淋清颗粒合用前后的肠吸收差异，从细胞、离体和在体不同角度证实了热淋清颗粒可显著降低左氧氟沙星的肠吸收，研究提示两药的相互作用可能主要产生于吸收阶段。并重点探讨了主要的代谢酶、转运体在介导其相互作用过程中的调控作用，采用RT-PCR和Western blot技术测定了肝脏中代谢酶（CYP2C9、CYP2D6、CYP3A4、CYP2C19、CYP2E1、CYP1A2）的表达，研究表明热淋清颗粒和左氧氟沙星合用后对肝脏代谢酶的影响较小；采用Western blot和免疫组化技术同时测定小肠和肾脏中转运蛋白（P-gp、MRP2、BCRP、OATP1A2）的表达，研究表明热淋清颗粒和左氧氟沙星合用后，可通过诱导外排转运蛋白使得两药合用后左氧氟沙星在体内的量显著降低，从转运体角度初步探讨了两药产生DDI的药代动力学机制。同时研究发现热淋清颗粒可降低尿路致病性大肠杆菌（UPEC）对膀胱上皮细胞的粘附，课题组将继续从UPEC菌毛和鞭毛入手，探讨热淋清颗粒及联合用药对细菌粘附、侵袭、运动的影响及相关粘附蛋白表达调控，为阐释热淋清颗粒临床应用科学内涵提供依据。