Zhang Biyu, Fu Denggang, Wang Xin, Hu Xuelei
Key Laboratory of Green Chemical Engineering Process of Ministry of Education, Hubei Key Laboratory of Novel Reactor and Green Chemical Technology, School of Chemical Engineering and Pharmacy, Wuhan Institute of Technology, Wuhan, China.
School of Medicine, Jiujiang University, Jiujiang, China.
Front Pharmacol. 2025 May 30;16:1591164. doi: 10.3389/fphar.2025.1591164. eCollection 2025.
Traditional Chinese medicine (TCM) has garnered attention for its potential in cancer therapy. Yinchen Wuling San (YWLS), a classical herbal formula, has been traditionally used for liver-related conditions, but its bioactive components and molecular mechanisms relevant to hematologic malignancies such as acute myeloid leukemia (AML) remain unclear. This study aims to identify the active compounds and potential molecular targets of Yinchen Wuling San in the context of AML through network pharmacology analysis, and to experimentally validate the effects of selected candidate compounds in AML models.
Active ingredients from six YWLS herbs were screened via the TCMSP database using oral bioavailability ≥30% and DL ≥0.18 thresholds. Targets were predicted using SwissTargetPrediction, and AML-related genes were obtained from DisGeNET and GeneCards. Key overlapping targets were analyzed via STRING PPI networks and GO/KEGG enrichment. Molecular docking was performed between three core compounds (genkwanin, isorhamnetin, quercetin) and hub proteins (e.g., SRC) using Sybyl-X. ADME profiles were predicted using SwissADME, and molecular dynamics simulations (GROMACS) assessed complex stability. These compounds were further evaluated (viability, apoptosis, cell cycle, RT-qPCR, flow cytometry) and using an AML xenograft mouse model.
Of 621 YWLS targets, 113 overlapped with 1,247 AML-related genes. PPI analysis identified hub genes, including AKT1, SRC, and EGFR. Enrichment analysis highlighted PI3K-AKT, MAPK, and JAK-STAT pathways. Genkwanin, isorhamnetin, and quercetin were predicted to target SRC, with strong molecular docking affinities. ADME analysis suggested favorable pharmacokinetics, and molecular dynamics simulations confirmed structural stability. , these compounds exhibited dose-dependent cytotoxicity, induced apoptosis, modulated the cell cycle, and downregulated SRC expression. Notably, Genkwanin promoted CD8 T cell proliferation and inhibited leukemia growth, improving survival in a leukemia xenograft model.
YWLS compounds, particularly Genkwanin, exhibit significant anti-leukemic activity via apoptosis induction, cell cycle modulation, and promote T cells proliferation. Genkwanin emerges as a promising therapeutic candidate for AML, warranting further clinical investigation.
中药在癌症治疗中的潜力已受到关注。茵陈五苓散（YWLS）作为一种经典的中药方剂，传统上用于治疗肝脏相关疾病，但其生物活性成分以及与急性髓系白血病（AML）等血液系统恶性肿瘤相关的分子机制仍不清楚。本研究旨在通过网络药理学分析确定茵陈五苓散在AML背景下的活性化合物和潜在分子靶点，并在AML模型中通过实验验证所选候选化合物的作用。
通过TCMSP数据库筛选茵陈五苓散六味药材中的活性成分，筛选标准为口服生物利用度≥30%且药物相似性（DL）≥0.18。使用SwissTargetPrediction预测靶点，并从DisGeNET和GeneCards获取AML相关基因。通过STRING蛋白质-蛋白质相互作用（PPI）网络以及基因本体（GO）/京都基因与基因组百科全书（KEGG）富集分析关键重叠靶点。使用Sybyl-X对三种核心化合物（芫花素、异鼠李素、槲皮素）与枢纽蛋白（如SRC）进行分子对接。使用SwissADME预测药物代谢动力学（ADME）特征，并通过分子动力学模拟（GROMACS）评估复合物稳定性。使用AML异种移植小鼠模型进一步评估这些化合物（活力、凋亡、细胞周期、实时定量聚合酶链反应（RT-qPCR）、流式细胞术）。
在621个茵陈五苓散靶点中，113个与1247个AML相关基因重叠。PPI分析确定了枢纽基因，包括蛋白激酶B1（AKT1）、SRC和表皮生长因子受体（EGFR）。富集分析突出了磷脂酰肌醇-3激酶-蛋白激酶B（PI3K-AKT）、丝裂原活化蛋白激酶（MAPK）和Janus激酶-信号转导及转录激活因子（JAK-STAT）信号通路。预测芫花素、异鼠李素和槲皮素靶向SRC，具有较强的分子对接亲和力。ADME分析表明其具有良好的药代动力学特征，分子动力学模拟证实了结构稳定性。这些化合物表现出剂量依赖性细胞毒性，诱导凋亡，调节细胞周期，并下调SRC表达。值得注意的是，芫花素促进CD8 T细胞增殖并抑制白血病生长，提高了白血病异种移植模型的生存率。
茵陈五苓散化合物，特别是芫花素，通过诱导凋亡、调节细胞周期以及促进T细胞增殖表现出显著的抗白血病活性。芫花素成为AML有前景的治疗候选药物，值得进一步临床研究。
