Network Pharmacology Based in Silico and in Vitro Anticancer Effects of Artemisia Maritima Extract in Human Prostate Cancer Cells

Artemisia maritima is an important medicinal plant with various bioactive molecules. Our study investigates the therapeutic potential of A. maritima extract against prostate cancer using a combination of network pharmacology, molecular docking, molecular dynamics, and in vitro methods. The phytochemicals of A. maritima and prostate cancer-related targets were retrieved from various databases followed by constructing a Protein-Protein Interaction (PPI) network. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were also conducted. Hub genes were used for molecular docking and dynamics to assess the interactions between target proteins and phytochemicals and their stability. The ethyl acetate extract of the aerial plant parts was prepared using Soxhlet method and then subjected to bioassays including MTT, clonogenic, and flowcytometry for toxicity (cytotoxicity and colony inhibition) and apoptotic assessments in prostate cancer in vitro. A total of 56 compounds were initially screened from the IMPPAT database, of which 22 were selected based on molecular weight ((≥500 Da)) drug-likeness, oral bioavailability (≥0.55, and gastrointestinal absorption (high/moderate). These compounds were further examined for interactions with prostate cancer-related targets from the GeneCards database, resulting in 66 shared targets. Network pharmacology identified hub genes such as NFKB1, MAPK1, and PIK3CD, which are involved in cancer progression pathways. GO and KEGG analyses highlighted these genes' roles in regulating cell death, intracellular signaling, and prostate cancer-specific pathways like PI3K-Akt and VEGF signaling. Molecular docking using CB-Dock2 revealed strong interactions between key compounds—Santamarine, Davanone, and Hydroxydavanone—and the hub genes, suggesting modulation of essential cancer pathways. Molecular dynamics simulations with iMODS and CABS-flex confirmed the stability of these protein-ligand complexes. In vitro experiments using LNCaP prostate cancer cells validated these findings, showing dose-dependent cytotoxicity in MTT assays (reducing viability to 20% and 10% after 24h (**p<0.01) and 48h (*p<0.05) of treatment, reduced colony formation in clonogenic assay to less than 50 (*p<0.05), apoptosis induction via Annexin V/PI staining. This is the first report on the anticancer potential of A. maritima against prostate cancer through network pharmacology, a significant plant in Traditional Chinese Medicine, offering insights into the molecular mechanisms.