The methanol extract from Cleistocalyx operculatus collected in Vietnam, was partitioned into hexane, ethyl acetate, and aqueous extract. A compounds was identified, including 2',6'-dihydroxy-4'-methoxychalcone (1), and guided by anti-inflammatory activity. Their chemical structures were elucidated by interpreting nuclear magnetic resonance (NMR) spectral data and comparing them with existing literature. The anti-inflammatory potential of isolated compounds was assessed by measuring their ability to inhibit nitric oxide (NO) production. Notably, 2',6'-dihydroxy-4'-methoxychalcone (1) significant NO production inhibition with an IC50 value of 47.58 ± 2.3 µM, comparable to the reference compound Positive control Cardamonin (3.02 µM) displayed little to no significant inhibitory activity and inhibited α-glucosidase, with an IC50 value of 161.3 µg/mL (the positive control acarbose, with an IC50 value of 214.1 µg/mL). These findings suggest that 2',6'-dihydroxy-4'-methoxychalcone from Cleistocalyx operculatus possess considerable potential as an inflammatory inhibitor and α-glucosidase further investigation and development.