The present study was designed to evaluate the therapeutic potential of bioactive compounds from chloroform extract of the leaves of Hylocereus undatus in the formation of advanced glycation end products (AGEs) in vitro. Bioactivity-guided fractionation of chloroform extract from Hylocereus undatus afforded two novel 12-ursen-type triterpenes, 3β, 16α, 23-trihydroxy-urs-12- en-28-oic acid ( 1) and 3β, 6β, 19α, 22α-tetrahydroxy-urs-12-en-28-oic acid ( 2), as well as four known triterpenes 2α, 3β, 23-tetrahydroxy-urs-11-en-28-oic acid ( 3), 3β-acetoxy-28-hydroxyolean-12-ene ( 4), 3β, 16α-dihidroxyolean-12-ene ( 5) and 3β-acetoxy-olean-12-ene ( 6). Our results revealed that triterpenes 1–3 were able to inhibit the formation of AGEs in all tested assays. The data indicated that the triterpenes had inhibitory activity at the múltiple stages of glycation and that there might be a high potential for decreasing protein oxidation and protein glycation that can enhance glycative stress in diabetic complications.