Dengue disease, which is caused by dengue virus (DENV) has been a major worldwide concern, with increased number of cases each year. Currently, there are no specific medications to treat the disease. Hence, there is a dire need to develop novel drugs for disease treatment. Glycolysis is a metabolic pathway that serves as the main source of energy for DENV replication and targeting the pathway is one of the ideal approach to discover new anti-DENV drugs. This paper focuses on the inhibition of human hexokinase isoform 2 (HK2) enzyme, which is one of the important enzymes in glycolysis, in the quest to disrupt DENV replication. In order to search for potential inhibitors, two methods were conducted, which are ligand-based screening and structure-based screening approaches. Docking of Daidzin , which was derived from Kudzu, a Japanese plant, into the active site of HK2 has shown the nearest binding affinity score (-7.94 kcal/mol) to glucose‘s (GLC), which is -8.15 kcal/mol. Meanwhile, Ethyl (2R)-2-[[3-[2-[(4-methylbenzoyl) amino]ethyl]-[1,2,4]triazolo[4,3-b]pyridazin-6-yl]sulfanyl]butanoate (Ethyl 2(R)), a compound which is the analogue of ((22E, 24R)-6β-methoxyergosta-7, 9(11), 22-triene-3β,5α-diol) or compound 2 from Ganoderma sinense obtained from ligand-based screening was also docked into the binding site of HK2, showing a binding affinity score of -8.43 kcal/mol. Both docking was conducted by using AutoDock4 software at virtual screening phase. These compounds were further analysed in an inhibition assay to determine the effects of these potential naturally-derived inhibitors on the activity of HK2. The outcome from the inhibition studies showed that both compounds exhibited substantial inhibition on the activity of HK2 enzyme, where Daidzin, at 0.5 mM, resulted in HK2 remaining activity of 87.28%, while Ethyl (2R) resulted in 70.09% of HK2 remaining activity at 0.5 mM concentration. The results also indicate that as the concentration of these compounds increased, the percentage of remaining enzyme activity decreased. In conclusion, this study has served as a platform for the development of anti-dengue drugs based on naturally-derived compounds, which is anticipated to be a safer option for dengue treatment.