| Summary: | <i>Nepenthes</i> are carnivorous pitcher plants that have several ethnobotanical uses, such as curing stomachache and fever. Here, we prepared different extracts from the stem, leaf, and pitcher of <i>Nepenthes miranda</i> to further investigate their pharmacological potential. The leaf extract of <i>N. miranda</i> obtained by 100% acetone (<i>N. miranda</i>-leaf-acetone) was used in this study to analyze the cytotoxic activities, antioxidation capacity, antibacterial activity, and allantoinase (ALLase) inhibitory effect of this plant. The cytotoxic effects of <i>N. miranda</i>-leaf-acetone on the survival, apoptosis, and migration of the cancer cell lines PC-9 pulmonary adenocarcinoma, B16F10 melanoma, and 4T1 mammary carcinoma cells were demonstrated. Based on collective data, the cytotoxic activities of <i>N. miranda</i>-leaf-acetone followed the order: B16F10 > 4T1 > PC-9 cells. In addition, the cytotoxic activities of <i>N. miranda</i>-leaf-acetone were synergistically enhanced when co-acting with the clinical anticancer drug 5-fluorouracil. <i>N. miranda</i>-leaf-acetone could also inhibit the activity of ALLase, a key enzyme in the catabolism pathway for purine degradation. Through gas chromatography–mass spectrometry, the 16 most abundant ingredients in <i>N. miranda</i>-leaf-acetone were identified. The top six compounds in <i>N. miranda</i>-leaf-acetone, namely, plumbagin, lupenone, palmitic acid, stigmast-5-en-3-ol, neophytadiene, and citraconic anhydride, were docked to ALLase, and their docking scores were compared. The docking results suggested plumbagin and stigmast-5-en-3-ol as potential inhibitors of ALLase. Overall, these results may indicate the pharmacological potential of <i>N. miranda</i> for further medical applications.
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