A herbal prescription in traditional Chinese medicine (TCM) has great complexity, with multiple components and multiple targets, making it extremely challenging to determine its bioactive compounds. Yinchenhao Tang (YCHT) has been extensively used for the treatment of jaundice disease. Although many studies have examined the efficacy and active ingredients of YCHT, there is still a lack of an in-depth systematic analysis of its effective components, mechanisms, and potential targets—especially one based on clinical patients. This study established an innovative strategy for discovering the potential targets and active compounds of YCHT based on an integrated clinical and animal experiment platform. The serum metabolic profiles and constituents of YCHT in vivo were determined by ultra-performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UPLC-Q-ToF-MS)-based metabolomics combined with a serum pharmacochemistry method. Moreover, a compound–target–pathway network was constructed and analyzed by network pharmacology and ingenuity pathway analysis (IPA). We found that eight active components could modulate five key targets. These key targets were further verified by enzyme-linked immunosorbent assay (ELISA), which indicated that YCHT exerts therapeutic effects by targeting cholesterol 7α-hydroxylase (CYP7A1), multidrug-resistance-associated protein 2 (ABCC2), multidrug-resistance-associated protein 3 (ABCC3), uridine diphosphate glucuronosyl transferase 1A1 (UGT1A1), and farnesoid X receptor (FXR), and by regulating metabolic pathways including primary bile acid biosynthesis, porphyrin and chlorophyll metabolism, and biliary secretion. Eight main effective compounds were discovered and correlated with the key targets and pathways. In this way, we demonstrate that this integrated strategy can be successfully applied for the effective discovery of the active compounds and therapeutic targets of an herbal prescription.

中药方剂复杂，成分多，靶点多，确定其生物活性成分极具挑战性。茵陈蒿汤（YCHT）已被广泛用于治疗黄疸病。尽管许多研究对YCHT的功效和活性成分进行了研究，但仍缺乏对其有效成分、机制和潜在靶点的深入系统分析——尤其是基于临床患者的分析。本研究建立了基于临床和动物实验综合平台的创新策略，用于发现YCHT的潜在靶点和活性化合物。YCHT在体内的血清代谢谱和成分通过基于超高效液相色谱-四极杆飞行时间质谱 (UPLC-Q-ToF-MS) 的代谢组学结合血清药物化学方法测定。此外，通过网络药理学和独创性途径分析（IPA）构建并分析了化合物-靶标-途径网络。我们发现八个活性成分可以调节五个关键目标。这些关键靶点通过酶联免疫吸附试验 (ELISA) 进一步验证，表明 YCHT 通过靶向胆固醇 7α-羟化酶 (CYP7A1)、多药耐药相关蛋白 2 (ABCC2)、多药耐药相关蛋白发挥治疗作用3 (ABCC3)、尿苷二磷酸葡萄糖醛酸转移酶 1A1 (UGT1A1) 和法尼醇 X 受体 (FXR)，并通过调节包括初级胆汁酸生物合成在内的代谢途径，卟啉和叶绿素代谢，胆汁分泌。发现了八种主要有效化合物，并与关键目标和途径相关联。通过这种方式，我们证明了这种综合策略可以成功应用于有效发现草药处方的活性化合物和治疗靶点。