The present study is aimed at evaluating the chemoprotective effect of Enicostemma littorale in DEN induced hepato-carcinogenesis in Sprague Dawley rats. The preliminary steps involved extraction, phytochemical investigation, HPTLC study and In vitro antioxidant activity using DPPH, ABTS. and Initiation of HCC was done by single i.p injection of DEN at a dose of 200mg/kg. The MEEL received treatment for 90 days after 14 days of development of HCC and continued for entire study period, whereas the other two group given normal saline, 5-flurouracil (20mg/kg ) i.p. The results showed that the injection DEN lead to the development of liver tumors in rats. Significant effect of serum biochemical parameter like SGOT, SGPT, ALP, UREA, TOTAL PROTEIN and tumor marker was observed with depletion of endogenous antioxidants SOD, CAT, GSH, there by leading to higher LPO. The result exhibited that MEEL treatment (Preventive) group offered excellent shielding against HCC and displayed all the parameter in near normal range with a maintained antioxidant enzyme system. The result obtained showed that extracts were found to containing phenols at a concentration of 70.25 mg/g and flavonoids 26.03mg/g. HPTLC Analysis showed presence of Quercetin, Mangiferin, Gallic acid, Catechin and sweroside. In the DPPH, ABTS, FRAP, TRAP radical scavenging assay MEEL has displayed the highest antiradical activity in both assays and was also comparable with the standard qucertein The present study reveal the efficacy of the MEEL to prevent malignancy induced by chemical carcinogen and the phytoconstituents responsible for activity.