Coumarin metabolites in Ocimum: chemical diversity, biosynthetic pathways, and network pharmacology-based prediction...

Ocimum, a widely used medicinal and culinary herb, is known for its health-promoting properties, yet most research has focused on its volatile oils, leaving the therapeutic potential and biosynthesis of its coumarins largely unexplored. Despite reports of coumarin bioactivity in other plants, a comprehensive understanding of coumarin diversity, pharmacology, and genetic regulation within Ocimum remains lacking-limiting both its breeding potential and pharmaceutical applications. Here, we applied an integrated multi-omics approach-including UPLC-MS/MS metabolomics, transcriptomics, network pharmacology, and molecular docking-to systematically profile coumarin metabolites, investigate their medicinal potential, and uncover their biosynthetic underpinnings across ten Ocimum accessions. A total of 59 coumarins were identified, 58 of which displayed significant variation among accessions, highlighting pronounced intra-genus metabolic diversity. Network pharmacology revealed that 35 bioactive coumarins potentially interact with 263 human targets, forming a rich metabolite-target-disease-pathway network. These coumarins were enriched in cancer- and inflammation-related KEGG pathways (e.g., PI3K-Akt, Ras/MAPK, endocrine resistance) and GO terms (e.g., tyrosine kinase activity), with hub compounds such as isodemethylwedelolactone, coumestrol, daphnetin, umbelliferone, and agrimonolide showing strong binding affinities to key proteins (e.g., EGFR, MAPK1, CCND1; docking scores ≤ - 6 kcal/mol). Coumarin biosynthetic pathways were reconstructed for several major compounds, revealing genotype-specific accumulation patterns and associated differentially expressed genes. These findings fill a critical knowledge gap regarding Ocimum coumarins by linking their metabolic diversity to pharmacological function and underlying gene expression. Our results lay the groundwork for breeding high-value Ocimum cultivars and developing coumarin-based functional foods or multi-target therapeutic agents, particularly for cancer and inflammatory diseases.