On the basis of our earlier reported studies, Cinnamomum tamala (CT), a well-known spice, was chosen for further investigations to evaluate its antitumor activity, to identify the bioactive compound present and to understand the probable mechanism/s of action. In the present studies, amongst different fractions of methanol extract of CT, the petroleum ether fraction [CT (A)] showed significant in vitro cytotoxic activity on cancer cell lines with no toxic effect on normal human peripheral blood mononuclear cells, mouse peritoneal macrophages and mouse spleen cells. In the fibrosarcoma-induced mouse tumor model, CT (A) significantly inhibited tumor growth, arrested the tumor metastasis to major organs and normalized the blood parameters of tumor induced mice. CT (A) also showed anti-angiogenic activity in chick embryo chorioallantoic membrane assay. The bioactive compounds likely to be present in the CT (A) fraction were identified by GC-MS analysis. After in silico molecular docking studies with probable targets, compounds from the CT (A) fraction were predicted to inhibit topoisomerase I enzyme. Thus, components of the petroleum ether fraction of methanol extract of CT are potential candidates for chemoprevention and cancer treatment and further studies are essential in order to confirm their mechanism of action.