Ursolic　acid　(UA)　is　a　naturally　bioactive　product　that　exhibits　potential　anticancer　effects.　The　relatively　safe　and　effective　molecule　intrigued　us　to　explore　a　way　to　further　improve　its　anti-cancer　activity　and　tumor-targeting　specificity.　In　the　present　study,　a　series　of　structural　modifications　of　UA　was　achieved,　which　resulted　in　significant　increase　in　growth　inhibition　on　various　cancer　cell　lines　with　minimal　effects　on　normal　cells.　The　leading　molecule　US597　(UA-4)　caused　depolarization　of　mitochondrial　membrane　potential,　cell　arrest　in　G0/G1　phase　and　apoptosis/necrosis　in　a　dose-dependent　manner.　Structural　docking　suggested　that　the　carbon　chains　of　the　modified　UA　derivatives　compete　strongly　with　glucose　for　binding　to　glucokinase,　the　key　glycolysis　enzyme　presumably　active　in　cancer　cells.　The　combination　of　2-deoxy-D-glucose　(2-DG)　and　UA-4　induced　cell　cycle　arrest　in　G2/M　phase,　promoted　caspase-dependent　cell　death,　reduced　hexokinase　activity,　aggravated　depletion　of　intracellular　ATP,　decreased　lactate　production　and　synergistically　inhibited　cancer　cell　growth　in　vitro　(HepG2)　and　in　vivo　(H22).　Collectively,　our　findings　suggest　that　the　structural　modification　enhances　efficacy　and　selectivity　of　UA,　and　the　combination　of　UA-4　with　2-DG　produces　synergistic　inhibition　on　hepatoma　cell　proliferation　by　dual　targeting　of　apoptosis　and　glycolysis.　©　2014,　Nature　Publishing　Group.　All　rights　reserved.