The alkaloid ingredients were considered to be responsible for the diverse pharmacological activities of the medicinal plant Tabernaemontana corymbosa (Roxb. ex Wall.). In the current finding, the systematic phytochemical investigation on T. corymbosa have been achieved. One new indole alkaloid tabercorympyline A (1) along with seven known indoles (2-8) were isolated from T. corymobsa. Their structures were elucidated by means of spectroscopic techniques and quantum chemical calculations. Tabercorympyline A (1) possessed the indole skeleton with rare N-containing nine membered ring. Chemical information network was used to comprehensively discover the clues for the glioma therapeutic leads from T. corymbosa alkaloids (TA). Inspired by chemical information network analysis, all the isolated compounds have been further validated their anti glioma activities in glioma cell line U251. Interestingly, the compounds 2, 3, 5, and 6 exhibited significant inhibitory effects on glioma cells in vitro. Molecular docking was ultimately used to indicate possible binding performance and mechanism between active compounds (2-3) and the core targets. This study sequentially assembled chemical information and network analysis, phytochemistry, molecular docking, and in vitro activity validation to comprehensively explore the effective compounds, related targets, and potential mechanisms of TA therapy for glioma.