| Insomnia, a prevalent sleep disorder, exhibits significant correlations with cardiovascular diseases, diabetes mellitus, and mental health disorders, severely compromising individual' quality of life. While existing pharmacological treatments demonstrate clinical efficacy, chronic administration often induces drug resistance and dependence, posing challenges for long-term management. Ganmai Dazao Decoction (GMDZD), a classical Chinese herbal formula, has shown substantial therapeutic effects in ameliorating insomnia symptoms. Nevertheless, its molecular mechanisms of action remain incompletely elucidated, particularly regarding multi-component interactions and signaling pathway modulations. Objective This study integrated network pharmacology, molecular docking, and molecular dynamics simulation to explore the active ingredients and core therapeutic targets of GMDZD in treating insomnia. The research systematically elucidated its potential molecular pharmacological mechanisms, including the modulation of inflammatory pathways, oxidative stress, and neurotransmitter regulation, thereby providing reference for clinical applications. Methods In November 2024, this study screened the active components and associated targets of GMDZD from the INPUT database, while insomnia-related transcriptomic datasets were acquired from the Gene Expression Omnibus (GEO) repository. Comparative expression profiling of insomnia versus controls was performed using GEO2R, yielding differentially expressed genes (DEGs). Shared targets between GMDZD and insomnia were identified by Venny. The protein-protein interaction (PPI) network was subsequently constructed via STRING database and Cytoscape 3.9.1. Shared targets underwent functional annotation through Gene Ontology (GO) and pathway enrichment analysis via the Kyoto Encyclopedia of Genes and Genomes (KEGG). The top three key active ingredients and top ten core targets were selected based on degree value. Molecular docking and molecular dynamics simulations were performed using AutoDock 4.4.6 to analyze the interactions between receptors and ligands. The stability of the receptor-ligand complexes was further validated, and the results were visualized using PyMOL 3.0.3. Results 337 active ingredients and 5,265 drug-related targets in GMDZD were found, and a total of 1,061 insomnia-associated differentially expressed genes (DEGs) were collected. Intersection analysis revealed 287 shared targets between GMDZD and insomnia. The herb-active ingredient-shared target-disease network highlighted quercetin, catechin, and kaempferol as key therapeutic components of GMDZD for insomnia. These three components exerted effects by targeting 10 core proteins: Nuclear Factor Kappa B Inhibitor Alpha (NFKBIA), Fibronectin 1 (FN1), Interleukin-6 (IL-6), FOS Proto-Oncogene (FOS), E1A Binding Protein P300 (EP300), Histone Deacetylase 1 (HDAC1), JUN Proto-Oncogene (JUN), Heat Shock Protein 90 Alpha Family Class A Member 1 (HSP90AA1), Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH), and Interleukin-1 Beta (IL-1β). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicated that GMDZD main likely alleviate insomnia by modulating the Interleukin-17 (IL-17) signaling pathway, lipid and atherosclerosis pathways. Molecular docking demonstrated strong binding affinities between the three key components and the 10 core targets. Molecular dynamic simulations showed that quercetin-GAPDH, catechin-HDAC1 and kaempferol-EP300 complexes could all reach a stable state. Conclusion The bioactive constituents of GMDZD—quercetin, catechin, and kaempferol—demonstrate multi-target therapeutic potential against insomnia by modulating ten core targets through key pathways such as IL-17 signaling pathway and lipid atherosclerosis pathway. |